[Histonet] BMP-2 protocol for bone

[Eddie Martin]

Hi Lindsay, 

There are a few complications  that will automatically come up due to working with rabbit tissue, and also due to the antibody from Abcam that you are using. 

I called Abcam prior to emailing you on a hunch that I had and was correct. The antibody you purchased from Abcam is unconjugated and first needs to be conjugated to whatever method you’re going to use. You could conjugate the antibody yourself. I already reached out to Abcam. It’s going to take them a few business days to get back to me to see whether their own antibody can be conjugated for IHC if that’s the route you’re going to go. 

Keeping this email simple for a response to your question, it may be easier to use another detection method for conjugating this specific antibody to rabbit bone...possibly FITC conjugated tyramide labeling with Alexa-Fluor. 

If you are insistent on using HRP it may be easier to contact me and we can further discuss as there are easily two complications that will arise due to the antibody you chose to optimize with and the species you are trying to stain on. 

Since you left your lab number on your post, I called and left my phone number on the voicemail for the young Lab if you would like to discuss and troubleshoot over the phone. I can also be reached via email at work at Eddie.martin at NIH.gov

Thanks,
Eddie

Eddie Martin HT(ASCP), QIHC(ASCP),HTL
Hematology Team Lead / Histology Technical Supervisor
The National Institutes of Health 
Clinic Center - Department of Laboratory Medicine
10 Center Drive, Room 2C 360
Bethesda, MD 20814



> On Oct 18, 2018, at 12:57 PM, Wilson, Lindsay A <Lindsay.A.Wilson at uth.tmc.edu> wrote:
> 
> Good morning,
> 
> Does anyone have a proven IHC protocol for BMP-2 on bone tissue? A big plus, if you have worked with it on rabbit bone!
> I am currently using Abcam’s BMP-2 (ab6285) with no success. I have performed antigen retrieval and non-antigen retrieval. I have used a dilution as low as 1:50. I appreciate any suggestions.
> 
> Lindsay Wilson, LVT, RLATG
> Young Laboratory
> 
> UTHealth | The University of Texas Health Science Center at Houston | School of Dentistry
> Department of Oral & Maxillofacial Surgery
> 
> 7500 Cambridge St. | Suite 6510 | Houston, TX 77054
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> 
>