Acetylcholinesterase staining.
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Last Update: 2020-10-22
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Source: Internet
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Author: User
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: I would like to use acetylcholinesterase dyeing mice colon smooth muscle choline energy nerve display, and use software for quantitative analysis, but I do not know the dyeing method used
reagents
and specific methods, shanghai where can do the dyeing? Please help, man!
I do not know this method with the intestinal mucosa after the plasma determination of acetylcholinesterase, which is better? Thank you!
answer: Acetylcholinesterase . Staining step
1, incubated in the following solution for 15 minutes 37C
iodized acetyl sulfur choline 12.5 mg
distilled water 2.5 ml
0. 82% sodium acetate 15.5 ml
0. 6% acetic acid 0.5 ml
2. 94% sodium citrate 1.5 ml
0. 75% copper sulfate 2.5 ml
0. 165% potassium iron cyanide 2.5 ml
(total 25 ml)
2, washing
3, sumu fine dyeing 1 minute
4, washing 1-2 minutes
5, dehydration, transparency, sealing
acetylcholinease dyeing Methods 2
staining steps
1, formarin before fixing 15 to 25 minutes
2, deionized water fully rinsed twice, each time 1 minute
3, incubated in the substation below for 4 hours, 40C
iodized acetyl sulfur choline 5 mg
0. 1M Sodium Malayate buffer PH6.0 6.5 ml
0. 1M sodium phosphate solution 0.5 ml
0. 03M copper sulfate 1 ml
distilled water 1 ml
0. 005M high-speed iron potassium cyanide 1 ml
asked: Thank you gao people for their pointing, I would like to ask where potassium iron cyanide can be bought ah,
organization
want to freeze treatment?
: Potassium iron cyanide, generally
chemical reagents
center can be purchased, our laboratory using frozen
slices
. It should be noted that acetylcholinesterase dyeing needs ionless water, when entering the reaction fluid to be washed clean with ionless water (3 times, 2min each time), the reaction fluid to be ready to use, can not exceed 30min, the pH value of the reaction fluid is also very important, otherwise it will affect the effect of brown-red precipitation.
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