Anti-BrdU antibody [IIB5] (ab8152)

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I just wanted to follow up on your query with some additional information.

For ab8039, this antibody can be used for detection of BrdU and BrU. We have not tested it regarding other chemicals of this family.

For ab8152, this antibody is specific for DNA in which BrdU has been incorporated. In immunoassays this antibody reacts strongly with free or carrier-protein coupled BrdU but not with other nucleosides. In immuncytochemistry the antibody only recognizes BrdU in denaturated (single stranded) DNA. The BrdU antibody is 100% crossreactive with Iodo-Deoxy-Uridine (IrdU). Therfore, IdU instead of BrdU can be used in studies.

I hope this information helps. Please contact us with any other questions.

Thank you for contacting us.

For floating sections, please check thefollowing protocol on IHC World website:
http://www.ihcworld.com/_protocols/epitope_retrieval/free_floating_sections.htm
This protocol is for paraffin-embedded as well as frozen sections.

As for ensuring the paraffin sections stick to your slides:
Different sections and tissues stick with differetn strength to the slides.
It might be advisable to use charged slides, as well as to bake your sections onto the slides by exposing them to heat (e.g. heating plate or oven) up to 60 degree Celsius.The purpose is to remove miscroscopic amounts of water between the sections and the slide.
Subsequently the paraffin would need to removed according to a standard IHC-P protocol. Then you would proceed with the antigen retrieval step.

Also, please check this information on the IHC World website for further advice.
http://www.ihcworld.com/_faq/histology-faq/section/s1.htm

The slides need to be immersed, for the retrieval to be effective. Yes, the sections could come off ifthey are kept too long in the retrieval solution. Should they come off, you would need to reduce the time they are exposed to the retrieval buffer.

Are the floating sections paraffin-embedded or frozen? I have found the following protocol on IHC World website which should help you further:
http://www.ihcworld.com/_protocols/epitope_retrieval/free_floating_sections.htm

I have checked with the lab:
Antigen retrieval can be done with HIER using citrate buffer.

Please see below for the protocol:

High Temperature antigen Unmasking Technique using Sodium Citrate Buffer for Immunohistochemical Demonstration on Paraffin Sections

1. Cut and mount sections on slides coated with Vectabond or APES (3-aminopropyltriethoxysilane).
2. Deparaffinize sections and rehydrate to distilled water.
3. Bring 1600ml 0.01M sodium citrate buffer (pH 6.0) to the boil in a Prestige stainless steel pressure cooker, using a hot plate. Cover but do not lock lid.
4. Position slides into metal staining racks and lower into pressure cooker ensuring slides are well immersed in citrate buffer. Lock lid. The small valve will rise.
5. When the pressure indicator valve (the large one) has risen after about 4 minutes, incubate sections for 1 minute.
6. Remove pressure cooker from heat source and run under cold water with lid on. When the small valve sinks open lid and remove slides and place immediately into distilled water. DO NOT OPEN LID UNTIL THE SMALL VALVE SINKS.
7. Wash sections in TBS buffer (pH 7.6) for 1 x 5 minutes.
8. Place sections in 1.5% hydrogen peroxide/methanol for 10 minutes.
9. Wash sections in distilled water for 2 x 5 minutes, then wash sections in TBS buffer for 2 x 5 minutes.
10. Place sections in normal serum for 20 minutes.
11. Cover sections with primary antibody. ( The optimal dilution of the antibody, incubation time and incubation temperature should be determined by the individual laboratory).
12. Wash in TBS buffer for 2 x 5 minutes.
13. Incubate sections in secondary antibody for 30 minutes.
14. Wash in TBS buffer for 2 x 5 minutes.
15. Incubate slides in ABComplex for 30 minutes.
16. Wash in TBS buffer for 2 x 5 minutes.
17. Incubate slides in DAB.
18. Wash in water for 2 x 5 minutes.
19. Counterstain with haematoxylin (if required), dehydrate, coverslip and mount.


To avoid sections becoming detached, sections should be mounted on Vectabond or APES covered slides, then dried at 37oC overnight followed by drying at 56oC for 60 minutes.

Thank you for contacting us with your question. We unfortunately do not have any data regarding the minimum length of ssDNA required for antibody staining on adherent cells.