Recommendations for antibodies for NHP FFPE tissues

[rosedever]

Hello,

I am working on a panel for multiplexed imaging of NHP FFPE lung and liver samples. I found these antibodies for human lung and liver through the OMAP initiative:

• ARG1: Cell Signaling Technology 93668S
• ALB: Abcam ab240330; clone: EPR12774
• ABCA3: Novus Biologicals NBP1-89310

Would these work in my tissue?

I am interested in antibodies for labeling endothelial cells in NHP FFPE tissues. Do you know why CD31 (clone WM59) is less effective on FFPE sections than other methods of preparing tissues?

[radtkea]

Thank you for your question! NHP FFPE is an exciting frontier.

Glad you are using the OMAP initiative to find liver and lung specific targets. I am not sure if these antibodies with work in NHP. Please see my comments below.

ARG1: Cell Signaling Technology 93668S: This antibody is validated for human tissues (vendor supplied images) and has reported reactivity for Human (H), Mouse (M), and Rat (R) but not Monkey (Mk). I would not recommend buying/testing this antibody in NHP tissues.
ALB: Abcam ab240330; clone: EPR12774 Same thing with this antibody. No reported reactivity for monkey.
ABCA3: Novus Biologicals NBP1-89310 Only reported for human reactivity.

Endothelial markers:
My colleagues have validated this CD31 antibody for NHP FFPE tissues. It's available in many different conjugates from Abcam.
This CD34 antibody has been reported reactivity in NHP. We have used it for human FFPE tissues.
We report this LYVE-1 antibody in NHP FFPE tissues.

Here is a GREAT resource for antibodies validated for CODEX imaging of NHP FFPE samples. The authors report what works and does not work based on IHC validation! It doesn't look like they have any endothelial markers.

This paper on IMC imaging of NHP FFPE samples might also be useful for you as you build out panels. No endothelial markers.

One final thought is the use of a lectin to label endothelial cells. @colinchu72 has used lectins for IBEX imaging of the human retina.

Hope this helps! Happy to chat more and would love to help you share your knowledge with the community, when ready. You can grab the latest reagent_resources.csv file from GitHub. Follow this template when recording your antibodies. Let me know if I can introduce you to any contacts with antibody suppliers. They are always looking for people to validate and report reactivity in NHP samples. Best of luck!

[radtkea]

I believe this is the lectin Colin used to label endothelial cells of the human retina.

[rosedever]

Those are fantastic resources, thank you @radtkea!

One resource that I have been using is the NHP Reagents Reactivity page. I can look up the antibody before I buy it, to see if there is reported reactivity in each species, which has been helpful for me!

[radtkea]

Happy to help!

Thank you very much for sending the NHP Reagents Reactivity link. It's a very useful resource. Best of luck and please keep us posted:)