ИТАР-ТАСС : На 104-м году жизни скончалась Нобелевский лауреат Рита Леви-Монтальчини:
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Sunday, December 30, 2012
Thursday, December 20, 2012
Wednesday, December 19, 2012
Tuesday, December 18, 2012
Нацбанк Беларуси отказался от обслуживания силовых структур: 
Таким образом, эти организации переводятся на обслуживание в коммерческие банки
Таким образом, эти организации переводятся на обслуживание в коммерческие банки
В Беларуси собирают деньги на новый детский хоспис и в тоже время подписывают контракт на поставку 4 российских учебно-боевых самолетов Як-130 в Беларусь
В Беларуси собирают деньги на новый детский хоспис:
ОО «Белорусский детский хоспис» проводит акцию по сбору средств на строительство в Боровлянах нового хосписа, сообщает БелаПАН.
ОО «Белорусский детский хоспис» проводит акцию по сбору средств на строительство в Боровлянах нового хосписа, сообщает БелаПАН.
Самая папулярная рэлігія: Хрысьціянства застаецца самай папулярнай рэлігіяй. Да такой высновы прыйшлі спэцыялісты Pew Research Center, што базуецца ў Вашынгтоне.
"Камволь" своих не сдает. Работники готовы защищать бывшего директора: 
Работники не отрицают, что зарплата и условия труда оставляют желать лучшего, но категорически не согласны с критикой в адрес бывшего директора.
Работники не отрицают, что зарплата и условия труда оставляют желать лучшего, но категорически не согласны с критикой в адрес бывшего директора.
Проблема "Беларусьфильма" — кинопрокат страны:
Одной из главных проблем белорусского кинематографа гендиректор "Беларусьфильма" Олег Сильванович назвал проблему выхода на внутренний рынок страны, пишет БелаПАН.
Одной из главных проблем белорусского кинематографа гендиректор "Беларусьфильма" Олег Сильванович назвал проблему выхода на внутренний рынок страны, пишет БелаПАН.
Жыхар Гродна: "На кухню ўжо не магу зайсці - такі холад": Канец свету ў Беларусі пачаўся з Гродна - горад застаўся без цяпла і паступова замярзае.
Белорусские коммунисты нашли своего идола даже под землей: Активисты белорусского общественного объединения "За Союз и коммунистическую партию Союза" обнаружили памятник Иосифу Сталину, который был демонтирован и закопан на территории республики в советские времена.
Как наехали на «Волата»?: Корыстные интересы российских властных элит могут разрушить наработанные десятилетиями кооперационные связи белорусских и российских оборонных предприятий и привести к утрате инновационного потенциала флагманов отечественной промышленности.
Ці нармальна гэта - забараняць звальняцца з працы?: Ці праглыне памяркоўны беларус новы дэкрэт, які фактычна ўводзіць прыгоннае права?
В Гродно на Архиерейском подворье освящен домовой храм: 
Храм освящен в честь особо почитаемого верующими святителя Николая Чудотворца...
Храм освящен в честь особо почитаемого верующими святителя Николая Чудотворца...
Monday, December 17, 2012
Here's A Great Primer On How Your Brain Works
Here's A Great Primer On How Your Brain Works:
If I had been asked 15 years ago to write a short piece about what the different parts of the brain did, it would have been a fairly straightforward task.
Not any more.
Over the last 15 years, the methods used to study the brain have advanced significantly, and with this so has our understanding.
Which makes the task of explaining the most complex organ in the body, well, complex.
Back to basics
The structural anatomy of the brain is certainly well defined and the more basic of our functions have been generally well mapped. The “lower levels,” such as the brainstem, regulate functions such as heart rate, breathing, and maintaining consciousness.
And the cerebellum is critical for the control and regulation of movement. While it was thought that this was its sole function, more recently the cerebellum has also been shown to have a role in so-called “higher functions” such as cognition and emotion.
As we move to the “higher levels” of the brain, namely the cerebral cortex, where more complex functions come into play, the assignment of function to structure becomes decidedly less distinct.
Different hemispheres
The cortex is structurally divided into two hemispheres (left and right) each with four lobes (occipital, parietal, temporal and frontal).
Brain functions, such as visual perception, language, memory, spatial ability and problem solving, have been traditionally allocated to one such lobe and/or hemisphere of the brain.
This has led to a number of misnomers regarding brain function, the most popular of which is the commonly held belief that there is a distinction between the left “logical” brain and the right “creative” brain. As discussed below, such complex behaviours are not determined by a specific brain region, or even a specific hemisphere.
The conceptualisation of an almost one-to-one relationship between structure and function was largely a result of lesion studies, where damage to a specific part of the brain resulted in impairments in a particular function. But as our techniques of assessing the brain became more sophisticated this approach was shown to be somewhat simplistic.
We have come a long way from the phrenology of Franz Gall in the 19th century, in which characteristics such as secretiveness, self-esteem and wonder were determined by the shape of the skull (thought to be a proxy of brain size), and the 20th century reliance on lesion studies to determine the function of the different areas of the brain.
Connected network
We are now developing an understanding that complex, higher-level brain functions are a result of a number of brain areas working together, in what are termed “networks”.
This has been a result of techniques such as Magnetic Resonance Imaging (MRI), which allows us to look at the entirety of brain regions involved in certain functions, with newer applications allowing the visualisation of connections between these brain regions (i.e. Diffusion Tensor Imaging).
This is not to say that there is no separation of function throughout the brain. Rather, while there are brain regions that carry out specialised functions, they are now thought to do so in concert with other brain regions via network connections.
To conceptualise this, you could think of the brain as a exceptionally efficient rail network where certain train stations perform specialised duties but they do so in conjunction with other stations, and they are connected and “communicate” via the rail network.
Language can provide a good example of how this occurs in the brain. Language is often thought of as a solely “left brain” function and, while there is a degree of lateralisation, this is certainly not the whole story.
There are specific regions in the dominant (usually left) hemisphere that are integral in the production and comprehension of speech, i.e. Broca’s area and Wernicke’s area respectively.
But the non-dominant (usually right) hemisphere is also involved in language, and is thought to be important in the recognition and production of the emotional context of speech.
Additionally, the “language network” involves a number of other dominant “left” hemisphere regions, including prefrontal cortex, premotor cortex, supplementary motor area, as well as regions of the parietal and temporal lobes.
These brain regions work together to perform higher order aspects of language such as the application of the correct syntax to speech, as well as the mapping of words to their meaning.
While there are certain highly-specialised brain regions for language, they are still part of an extensive network of brain regions which work together to produce this complex function.
In addition, the brain is not fixed in its functioning. It is plastic and, if needed due to illness or injury, it can recruit new regions and/or networks to take over the functions of the damaged areas.
And so we believe it is a complex interaction between structure and function that best describes what the different part of the brain do – at least for now …
Please follow Science on Twitter and Facebook.
Join the conversation about this story »
If I had been asked 15 years ago to write a short piece about what the different parts of the brain did, it would have been a fairly straightforward task.
Not any more.
Over the last 15 years, the methods used to study the brain have advanced significantly, and with this so has our understanding.
Which makes the task of explaining the most complex organ in the body, well, complex.
Back to basics
The structural anatomy of the brain is certainly well defined and the more basic of our functions have been generally well mapped. The “lower levels,” such as the brainstem, regulate functions such as heart rate, breathing, and maintaining consciousness.
And the cerebellum is critical for the control and regulation of movement. While it was thought that this was its sole function, more recently the cerebellum has also been shown to have a role in so-called “higher functions” such as cognition and emotion.
As we move to the “higher levels” of the brain, namely the cerebral cortex, where more complex functions come into play, the assignment of function to structure becomes decidedly less distinct.
Different hemispheres
The cortex is structurally divided into two hemispheres (left and right) each with four lobes (occipital, parietal, temporal and frontal).
Brain functions, such as visual perception, language, memory, spatial ability and problem solving, have been traditionally allocated to one such lobe and/or hemisphere of the brain.
This has led to a number of misnomers regarding brain function, the most popular of which is the commonly held belief that there is a distinction between the left “logical” brain and the right “creative” brain. As discussed below, such complex behaviours are not determined by a specific brain region, or even a specific hemisphere.
The conceptualisation of an almost one-to-one relationship between structure and function was largely a result of lesion studies, where damage to a specific part of the brain resulted in impairments in a particular function. But as our techniques of assessing the brain became more sophisticated this approach was shown to be somewhat simplistic.
We have come a long way from the phrenology of Franz Gall in the 19th century, in which characteristics such as secretiveness, self-esteem and wonder were determined by the shape of the skull (thought to be a proxy of brain size), and the 20th century reliance on lesion studies to determine the function of the different areas of the brain.
Connected network
We are now developing an understanding that complex, higher-level brain functions are a result of a number of brain areas working together, in what are termed “networks”.
This has been a result of techniques such as Magnetic Resonance Imaging (MRI), which allows us to look at the entirety of brain regions involved in certain functions, with newer applications allowing the visualisation of connections between these brain regions (i.e. Diffusion Tensor Imaging).
This is not to say that there is no separation of function throughout the brain. Rather, while there are brain regions that carry out specialised functions, they are now thought to do so in concert with other brain regions via network connections.
To conceptualise this, you could think of the brain as a exceptionally efficient rail network where certain train stations perform specialised duties but they do so in conjunction with other stations, and they are connected and “communicate” via the rail network.
Language can provide a good example of how this occurs in the brain. Language is often thought of as a solely “left brain” function and, while there is a degree of lateralisation, this is certainly not the whole story.
There are specific regions in the dominant (usually left) hemisphere that are integral in the production and comprehension of speech, i.e. Broca’s area and Wernicke’s area respectively.
But the non-dominant (usually right) hemisphere is also involved in language, and is thought to be important in the recognition and production of the emotional context of speech.
Additionally, the “language network” involves a number of other dominant “left” hemisphere regions, including prefrontal cortex, premotor cortex, supplementary motor area, as well as regions of the parietal and temporal lobes.
These brain regions work together to perform higher order aspects of language such as the application of the correct syntax to speech, as well as the mapping of words to their meaning.
While there are certain highly-specialised brain regions for language, they are still part of an extensive network of brain regions which work together to produce this complex function.
In addition, the brain is not fixed in its functioning. It is plastic and, if needed due to illness or injury, it can recruit new regions and/or networks to take over the functions of the damaged areas.
And so we believe it is a complex interaction between structure and function that best describes what the different part of the brain do – at least for now …
Please follow Science on Twitter and Facebook.
Join the conversation about this story »
The biotin-streptavidin interaction can be reversibly broken using water at elevated temperatures - Holmberg - 2005 - ELECTROPHORESIS - Wiley Online Library
Tuesday, August 28, 2012
Conquering Pain and Infection with Drugs from Nature’s Medicine Cabinet
http://www.faseb.org/Portals/0/PDFs/opa/natural%20products%20article%20FINAL.pdf
http://www.faseb.org/Policy-and-Government-Affairs/Publications/Breakthroughs-in-Bioscience.aspx
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http://www.faseb.org/Policy-and-Government-Affairs/Publications/Breakthroughs-in-Bioscience.aspx
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Magic Bullets and Monoclonals: An Antibody Tale
http://www.faseb.org/LinkClick.aspx?fileticket=qwHBXHJIMuY%3d&tabid=366
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Tuesday, August 21, 2012
Monday, August 20, 2012
Sunday, August 19, 2012
Saturday, August 18, 2012
Thursday, August 16, 2012
Минчане: Время Лукашенко прошло: Лукашенко побил личный рекорд правления Брежнева. Сумеет ли нынешний руководитель Беларуси обставить других известных диктаторов. Например Мао Цзе Дуна или Муаммара Каддафи?
Зенон Позняк: плюшевая акция принесла белорусам только вред: «В результате пустопорожней шведской провокации Лукашенко сел на коня своей антибелорусской политики, воспользовался случаем полностью»...
Польский шпион написал книгу о белорусском КГБ: Офицер польской разведки под псевдонимом Винсент В. Северский (Vincent V. Severski) написал шпионскую повесть, события которой разворачиваются в лукашенковской Беларуси и — сюрприз — ...в Швеции.
Дмитрий Усс: Я не виновен: "Незаконно осужденный" бывший кандидат в президенты Дмитрий Усс направил на имя председателя Верховного суда Валентина Сукало надзорную жалобу на приговор, вынесенный по событиям 19 декабря 2010 года. Это уже третья по счету надзорная жалоба.
Выйшаў першы нумар электроннага часопіса аб правах чалавека ў Беларусі: У першым нумары часопіса аўтары сфакусавалі сваю ўвагу на праблемах свабоды сходаў і справядлівага суда.
Беларусь заняла 91-е место из 145 в мировом "рейтинге здоровья": На строку выше нас расположилась Индонезия (33,10), ниже - Монголия (32,35). Россия - на 97-м месте с 26,44 балла.
Дамова з Ітэрай пра «Мінск-сіці» скасоўваецца: Інвэстыцыйная дамова паміж Рэспублікай Беларусь і сумесным таварыствам з абмежаванай адказнасьцю «Мінск-Сіці» аб будаўніцтве шматфункцыянальнага цэнтру «Дзелавы цэнтар «Мінск-Сіці» скасоўваецца.
Спортсмены считают, что на них не стоит давить: Очередные белорусские олимпийцы высказались против ультимативных медальных задач, которые ставило перед ними руководство…
Офицерам белорусской армии рекомендуют одеваться в секонд-хенде: Также было бы неплохо, если бы военнослужащие отказались от Интернета. Читайте трагикомедию о жизни Вооруженных Сил Беларуси в четырех актах.
Колькі яшчэ засталося Лукашэнку?: Пабіў асабісты рэкорд Брэжнева. Але ці паб'е цяперашні кіраўнік Беларусі рэкорды бесперапыннага знаходжання пры ўладзе такіх дыктатараў як Нурсултан Назарбаеў, Маў Цзэ Дун ці Муамар Кадафі?
Легко ли студенту снять квартиру в Гродно?: 
Доступным съемное жилье для студента не назовешь. Средняя цена за однокомнатную квартиру в Гродно – 110-120 $ без учета коммунальных платежей. И такое предложение еще нужно умудриться отыскать...
Доступным съемное жилье для студента не назовешь. Средняя цена за однокомнатную квартиру в Гродно – 110-120 $ без учета коммунальных платежей. И такое предложение еще нужно умудриться отыскать...
Wednesday, August 15, 2012
β Parenchyma and the Clearance of Interstitial Solutes, Including Amyloid A Paravascular Pathway Facilitates CSF Flow Through the Brain
http://stm.sciencemag.org/content/4/147/147ra111.full.pdf'
Wednesday, August 1, 2012
ScienceDirect.com - Journal of Immunological Methods - An improved method to create nitrocellulose particles suitable for the immobilization of antigen and antibody
Use of nitrocellulose films for affinity-directed mass spectrometry
http://onlinelibrary.wiley.com/store/10.1002/rcm.432/asset/432_ftp.pdf?v=1&t=h5cixgg9&s=66b49a79d3055e4405a8a004f6d7290187ca8a86&systemMessage=Wiley+Online+Library+will+be+disrupted+on+4+August+from+10%3A00-12%3A00+BST+%2805%3A00-07%3A00+EDT%29+for+essential+maintenance
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Tuesday, July 31, 2012
Antibodies bound to nitrocellulose in acidic buffers retain biological activity
http://onlinelibrary.wiley.com/store/10.1002/elps.11501401141/asset/11501401141_ftp.pdf;jsessionid=7EE206CC5B355F91DED6639A2533A160.d01t02?v=1&t=h5bgksjq&s=f5a7bb39b5444b2a1b4c276c516c9e3a9afbdcbf&systemMessage=Wiley+Online+Library+will+be+disrupted+on+4+August+from+10%3A00-12%3A00+BST+%2805%3A00-07%3A00+EDT%29+for+essential+maintenance
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Sunday, July 29, 2012
Saturday, July 28, 2012
Saturday, July 21, 2012
Международных наблюдателей предлагают послать на четыре буквы | БЕЛОРУССКИЕ НОВОСТИ: Лебедько предложил БДИПЧ ОБСЕ взять на себя иную функцию: создать рабочую группу или организовать круглый стол, перед которыми бы стояла задача отрегулировать белорусское избирательное законодательство и практику проведения выборов в соответствии с мировыми стандартами
Читать полностью:http://naviny.by/rubrics/elections/2012/07/21/ic_articles_623_178545/
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Читать полностью:http://naviny.by/rubrics/elections/2012/07/21/ic_articles_623_178545/
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Amyloid precursor protein promotes endoplasmic reticulum stress-induced cell death via C/EBP homologous protein-mediated pathway - Takahashi - 2009 - Journal of Neurochemistry - Wiley Online Library
The amyloid precursor protein protects PC12 cells against endoplasmic reticulum stress-induced apoptosis - Kögel - 2003 - Journal of Neurochemistry - Wiley Online Library
Tuesday, July 17, 2012
Immunoprecipitation (IP) technical guide and protocols
http://www.piercenet.com/files/TR0064-Immunoprecipitation-guide.pdf
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'via Blog this'
Thursday, July 12, 2012
A mutation in APP protects against Alzheimer/'s disease and age-related cognitive decline : Nature : Nature Publishing Group
A mutation in APP protects against Alzheimer/'s disease and age-related cognitive decline : Nature : Nature Publishing Group:
By highlighting the association between decreased APP processing and protection against Alzheimer’s disease, the research strongly supports the amyloid hypothesis, which has suffered recently, said Goate. “A lot of ongoing therapeutic work based on the amyloid hypothesis has failed,” she explained, “so people were worried that modulating amyloid might not work.” But the current study suggests that reducing amyloid β may play a role in preventing Alzheimer’s development. In addition, the data also support continued investigation of BACE inhibitors as possible therapeutic drugs to treat Alzheimer’s, she added
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By highlighting the association between decreased APP processing and protection against Alzheimer’s disease, the research strongly supports the amyloid hypothesis, which has suffered recently, said Goate. “A lot of ongoing therapeutic work based on the amyloid hypothesis has failed,” she explained, “so people were worried that modulating amyloid might not work.” But the current study suggests that reducing amyloid β may play a role in preventing Alzheimer’s development. In addition, the data also support continued investigation of BACE inhibitors as possible therapeutic drugs to treat Alzheimer’s, she added
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Wednesday, May 23, 2012
Tuesday, May 22, 2012
Sunday, May 20, 2012
Thursday, May 3, 2012
Tuesday, May 1, 2012
Nitrocellulose (Biorad Laboratories, Hercules, CA) was suspended
http://www.biomedcentral.com/content/pdf/1754-1611-1-2.pdf
Internal amino acid sequence analysis of proteins separated by oneor two-dimensional gel electrophoresis after in situ protease digestion on nitrocellulose
http://www.pnas.org/content/84/20/6970.full.pdf
'via Blog this'
'via Blog this'
Monday, April 30, 2012
«Разлаваў хам шклоўскага пана»: Новы альбом «Ляпіса Трубяцкога»: «Рабкор» з'явіўся ў інтэрнэце на Першамай.
Thursday, April 26, 2012
Wednesday, April 25, 2012
Saturday, March 31, 2012
Monday, March 19, 2012
Identification of membrane proteins from mammalian cell/tissue using methanol-facilitated solubilization and tryptic digestion coupled with 2D-LC-MS/MS : Article : Nature Protocols
Use of Nitrocellulose Membranes for Protein Characterization by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry
We present an improved method for MALDI-MS analysis
of proteins that have been electroblotted onto a nitrocellulose (NC) membrane. With this approach, electroblotted
proteins can be analyzed directly for intact molecular
weight determination or after on-membrane digestion by
dissolution of the nitrocellulose in MALDI matrix solution
containing 70% acetonitrile and 30% methanol. This
solution helps maintain solubility of proteins and peptides
while dissolving the NC membrane, which is dissolved
by 100% acetone in other protocols. On-membrane tryptic
digestion using this method requires half the time of ingel digestion and results in fewer missed cleavages and
better protein coverage. For the membrane proteins
studied, bovine uroplakins II and III, the protein coverage
was almost twice that provided by conventional in-gel
digestion, and the transmembrane domains of both
uroplakins were detected only after on-membrane digestion. We also demonstrated the compatibility with MALDIMS of a new dye, MemCode, which is specifically designed
for staining NC membrane-immobilized proteins and is
faster and more sensitive than Ponceau-S. Our improved
on-membrane digestion protocol greatly improves the
study of soluble and, particularly strikingly, integral
membrane proteins by mass spectrometry http://webdoc.nyumc.org/nyumc/files/sun-lab/attachments/2006.AC.MSa.pdf
'via Blog this'
of proteins that have been electroblotted onto a nitrocellulose (NC) membrane. With this approach, electroblotted
proteins can be analyzed directly for intact molecular
weight determination or after on-membrane digestion by
dissolution of the nitrocellulose in MALDI matrix solution
containing 70% acetonitrile and 30% methanol. This
solution helps maintain solubility of proteins and peptides
while dissolving the NC membrane, which is dissolved
by 100% acetone in other protocols. On-membrane tryptic
digestion using this method requires half the time of ingel digestion and results in fewer missed cleavages and
better protein coverage. For the membrane proteins
studied, bovine uroplakins II and III, the protein coverage
was almost twice that provided by conventional in-gel
digestion, and the transmembrane domains of both
uroplakins were detected only after on-membrane digestion. We also demonstrated the compatibility with MALDIMS of a new dye, MemCode, which is specifically designed
for staining NC membrane-immobilized proteins and is
faster and more sensitive than Ponceau-S. Our improved
on-membrane digestion protocol greatly improves the
study of soluble and, particularly strikingly, integral
membrane proteins by mass spectrometry http://webdoc.nyumc.org/nyumc/files/sun-lab/attachments/2006.AC.MSa.pdf
'via Blog this'
Mass spectrometric characterization of proteins transferred from polyacrylamide gels to membrane filters - Ino - 2011 - FEBS Journal - Wiley Online Library
У сацыяльных сетках выберуць лепшую беларускую прозу: Агенцтва рэгіянальнага развіцця “Дзедзіч” у межах кампаніі “Наша мова” адкрывае галасаванне за найлепшую прозу на беларускай мове.
больш па тэме
Аудио:
Analysis of Electroblotted Proteins by Mass Spectrometry: Protein Identification after Western Blotting
We describe a new approach for the identification and characterization by mass spectrometry of
proteins that have been electroblotted onto nitrocellulose. Using this method (Blotting And
Removal of Nitrocellulose, or BARN), proteins can be analyzed either as intact proteins for
molecular weight determination or as peptides generated by on-membrane proteolysis. Acetone is
used to dissolve the nitrocellulose and to precipitate the adsorbed proteins/peptides, thus
removing the nitrocellulose which can interfere with mass spectrometry analysis. This method
offers improved protein coverage, especially for membrane proteins such as uroplakins, since the
extraction step after in-gel digestion is avoided. Moreover, removal of nitrocellulose from the
sample solution allows sample analysis by both MALDI- and (LC) ESI-based mass
spectrometers. Finally, we demonstrate the utility of BARN for the direct identification of soluble
and membrane proteins after Western blotting, obtaining comparable or better results than with
in-gel digestion.
http://www.mcponline.org/content/early/2007/10/15/mcp.M700415-MCP200.full.pdf
proteins that have been electroblotted onto nitrocellulose. Using this method (Blotting And
Removal of Nitrocellulose, or BARN), proteins can be analyzed either as intact proteins for
molecular weight determination or as peptides generated by on-membrane proteolysis. Acetone is
used to dissolve the nitrocellulose and to precipitate the adsorbed proteins/peptides, thus
removing the nitrocellulose which can interfere with mass spectrometry analysis. This method
offers improved protein coverage, especially for membrane proteins such as uroplakins, since the
extraction step after in-gel digestion is avoided. Moreover, removal of nitrocellulose from the
sample solution allows sample analysis by both MALDI- and (LC) ESI-based mass
spectrometers. Finally, we demonstrate the utility of BARN for the direct identification of soluble
and membrane proteins after Western blotting, obtaining comparable or better results than with
in-gel digestion.
http://www.mcponline.org/content/early/2007/10/15/mcp.M700415-MCP200.full.pdf
Internal amino acid sequence analysis of proteins separated by oneor two-dimensional gel electrophoresis after in situ protease digestion on nitrocellulose
http://www.pnas.org/content/84/20/6970.full.pdf
We have developed a general two-step method for obtaining peptide fragments for sequence analysis from
picomole quantities of proteins separated by gel electrophoresis. After separation by one- or two-dimensional polyacrylamide gel electrophoresis, proteins are electrophoretically transferred (electroblotted) onto nitrocellulose, the protein-containing regions are detected by reversible staining and are cut out,
and each protein is digested in situ by proteolytic enzymes such
as trypsin or staphylococcal V-8 protease. The resulting peptide
fragments are separated by narrow-bore reverse-phase HPLC,
collected, and sequenced in a gas-phase sequenator. Excellent
peptide recoveries and the absence of extraneous contaminants
in the separation of the peptide fragment mixture allow the
generation of extensive internal sequence information from
picomole amounts of protein. The method thus overcomes the
problem of obtaining amino acid sequence data from Nterminally blocked proteins and provides multiple, independent stretches of sequence that can be used to generate
oligonucleotide probes for molecular cloning and/or used to
search sequence data bases for related proteins. This method
has been successfully applied to the routine amino acid sequence analysis of a wide range of proteins isolated from oneand two-dimensional polyacrylamide gels.
'via Blog this'
We have developed a general two-step method for obtaining peptide fragments for sequence analysis from
picomole quantities of proteins separated by gel electrophoresis. After separation by one- or two-dimensional polyacrylamide gel electrophoresis, proteins are electrophoretically transferred (electroblotted) onto nitrocellulose, the protein-containing regions are detected by reversible staining and are cut out,
and each protein is digested in situ by proteolytic enzymes such
as trypsin or staphylococcal V-8 protease. The resulting peptide
fragments are separated by narrow-bore reverse-phase HPLC,
collected, and sequenced in a gas-phase sequenator. Excellent
peptide recoveries and the absence of extraneous contaminants
in the separation of the peptide fragment mixture allow the
generation of extensive internal sequence information from
picomole amounts of protein. The method thus overcomes the
problem of obtaining amino acid sequence data from Nterminally blocked proteins and provides multiple, independent stretches of sequence that can be used to generate
oligonucleotide probes for molecular cloning and/or used to
search sequence data bases for related proteins. This method
has been successfully applied to the routine amino acid sequence analysis of a wide range of proteins isolated from oneand two-dimensional polyacrylamide gels.
'via Blog this'
On-Membrane Tryptic Digestion of Proteins for Mass Spectrometry Analysis
Identification of proteins and characterization of posttranslational modifications are crucial steps for
many biological, biochemical, and biomedical studies, and mass spectrometry has become the method of
choice for these analyses. Here we describe two methods for the on-membrane digestion of proteins electroblotted onto nitrocellulose membranes prior to analysis by mass spectrometry. These on-membrane
methods take approximately half the time of in-gel digestion and provide better digestion efficiency, due
to the better accessibility of the protease to the proteins adsorbed onto the nitrocellulose, and better
protein sequence coverage, especially for membrane proteins where large and hydrophobic peptides are
commonly present.
http://www.springerlink.com/content/l227u63572311256/#section=55463&page=1
http://www.springerlink.com/content/l227u63572311256/fulltext.pdf
many biological, biochemical, and biomedical studies, and mass spectrometry has become the method of
choice for these analyses. Here we describe two methods for the on-membrane digestion of proteins electroblotted onto nitrocellulose membranes prior to analysis by mass spectrometry. These on-membrane
methods take approximately half the time of in-gel digestion and provide better digestion efficiency, due
to the better accessibility of the protease to the proteins adsorbed onto the nitrocellulose, and better
protein sequence coverage, especially for membrane proteins where large and hydrophobic peptides are
commonly present.
http://www.springerlink.com/content/l227u63572311256/#section=55463&page=1
http://www.springerlink.com/content/l227u63572311256/fulltext.pdf
Sunday, March 18, 2012
Are amyloid-degrading enzymes viable therapeutic targets in Alzheimer’s disease? - Nalivaeva - 2011 - Journal of Neurochemistry - Wiley Online Library
Aminopeptidase A contributes to the N-terminal truncation of amyloid β-peptide - Sevalle - 2009 - Journal of Neurochemistry - Wiley Online Library
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