Biology版 - 问一下methanol/acetic acid的fixation |
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s******s
发帖数: 13035 | 1 要研究DNA/蛋白作用啥的，somehow后面的步骤不太适合formaldehyde，
打算用甲醇/乙酸固定，然后做pull down一类的，请问一下这个可行么？
谁能指教一下这个fix的原理是啥，有covalent bond么？ | T**********t
发帖数: 1604 | 2 主要是让蛋白变性沉积，跟covalent bond没啥关系。
http://en.wikipedia.org/wiki/Fixation_%28histology%29
Precipitating fixatives - Alcohols
Precipitating (or denaturing) fixatives act by reducing the solubility of
protein molecules and (often) by disrupting the hydrophobic interactions
that give many proteins their tertiary structure. The precipitation and
aggregation of proteins is a very different process from the crosslinking
that occurs with the aldehyde fixatives.
The most common precipitating fixatives are ethanol and methanol. They are
commonly used to fix frozen sections and smears. Acetone is also used and
has been shown to produce better histological preservation than frozen
sections when employed in the Acetone Methylbenzoate Xylene (AMEX) technique.
The protein denaturants - methanol, ethanol and acetone - are rarely used
alone for fixing blocks unless studying nucleic acids.
Acetic acid is a denaturant that is sometimes used in combination with the
other precipitating fixatives. The alcohols, by themselves, are known to
cause considerable shrinkage and hardening of tissue during fixation while
acetic acid alone is associated with tissue swelling; combining the two may
result in better preservation of tissue morphology. | s******s
发帖数: 13035 | 3 太感谢了！！我咋就wiki不到呢。
【在 T**********t 的大作中提到】
: 主要是让蛋白变性沉积，跟covalent bond没啥关系。
: http://en.wikipedia.org/wiki/Fixation_%28histology%29
: Precipitating fixatives - Alcohols
: Precipitating (or denaturing) fixatives act by reducing the solubility of
: protein molecules and (often) by disrupting the hydrophobic interactions
: that give many proteins their tertiary structure. The precipitation and
: aggregation of proteins is a very different process from the crosslinking
: that occurs with the aldehyde fixatives.
: The most common precipitating fixatives are ethanol and methanol. They are
: commonly used to fix frozen sections and smears. Acetone is also used and
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