 Home > Biochemistry News > Biotechnology News > T-cell dependent antigen specific in situ staining (Pe)

T-cell dependent antigen specific in situ staining (Pe)

 Last Update: 2020-10-26
 Source: Internet
 Author: User

Double staining for Ag and Ki-67, TUNEL or BCL-6.

Cut frozen tissue sections (spleen, lymph nodes) in a cryostat at~ 4µm, on plain glass slides (Adhesive -coated slides are sometimes too hydrophobic, but fine if you have them).

1- Fix the sections in 10% buffered formalin for 10 min. at RT.

10- add the HRP-conjugated Goat anti rabbit (Vector 50 to 100 µl, dilution 1:100) and incubate for 30’.

Use tyramide amplification here
Continue
Biotin-conjugated secondary Ab choice:
10- add the biotin-conjugated goat anti-rabbit (50 to 100 µl, dilution 1:200) and incubate for 30 min.

12 -add the HRP-conjugated avidin (50 to 100 µl, dilution 1:300 -500) and incubate for 20 min. Be careful not to dilute the avidin in biotin-containing medium.

11 -fix in absolute methanol RT for 10 min. Warning: methanol dissolves AEC and blocks HRP. Do this step only after biotin-secondary or tyramide-biotin.

19- apply the diluted antibody (BLC-6, Ki-67, others)
20- wash twice in TBS-T
21- counterstain with an AP-conjugated secondary antib

This article is an English version of an article which is originally in the Chinese language on echemi.com and is provided for information purposes only. This website makes no representation or warranty of any kind, either expressed or implied, as to the accuracy, completeness ownership or reliability of the article or any translations thereof. If you have any concerns or complaints relating to the article, please send an email, providing a detailed description of the concern or complaint, to service@echemi.com. A staff member will contact you within 5 working days. Once verified, infringing content will be removed immediately.

Related Products

L-Ascorbate-2-Monophosphate Feed Grade

CAS No.: 109620-90-8

Feed Grade

Potassium Sorbate from reliable manufacturer

CAS No.: 590-00-1

Food Grade

Doxycycline Hydrochloride

CAS No.: 10592-13-9

Pharmaceutical Grade

Hot Tags

united states jobs(223)

jobs united states(236)

1 year treasury(277)

1 year subscription(296)

announcements today(81)

80 20 company(94)

united states logo(227)

single use technology(195)

healthy food name(42)

ups united states(221)

Hot Articles

The source of this page with content of products and services is from Internet, which doesn't represent ECHEMI's opinion. If you have any queries, please write to service@echemi.com. It will be replied within 5 days.

Moreover, if you find any instances of plagiarism from the page, please send email to service@echemi.com with relevant evidence.