I am new to immune oncology. What are the best antibodies against CTLA-4, Tigit, Lag-3 and Tim-3 for IHC and Flow cytometry? Are there any problems I need to be aware of?
Dear Martin Schmidt In our lab, we were using BD BV421 CD152
(Cat. No. 562743 ) for human samples.
But that depends on the specificity of available cytometer, you may choose any other fluorochrome as well. For us, this product worked well.
Now i am thinking to shift this to blue laser line fluorochrome (PE or FITC).
Hello Martin!
Are you looking for anti-mouse or anti-human antibodies?
For mouse I can recommend the following antibody clones for flow cytometry:
LAG-3: C9B7W
TIM-3: RTM3-23
TIGIT: RMT3-23
CTLA-4: UC10-4B9 is what we use in our lab, but I never used it myself
Depending on your setting and staining protocol, I found that using fairly high concentrations of these antibodies is necessary compared to other markers like CD4/ CD8, CD44,... (In the range of 1-2µL/100µL staining solution for ~1E6 cells) to get a good signal. You should titrate them for optimal resolution. Depending on your model/ experiment, there is a high chance of co-expression of these markers, so keep that in mind during panel design (i.e. try to use colors with little spectral overlap for these markers if possible).
As for IHC, I don´t have any experience with these clones, but I guess they should work just as well.
Dear Martin Schmidt In our lab, we were using BD BV421 CD152
(Cat. No. 562743 ) for human samples.
But that depends on the specificity of available cytometer, you may choose any other fluorochrome as well. For us, this product worked well.
Now i am thinking to shift this to blue laser line fluorochrome (PE or FITC).
Thank you for the helpful recommendations! We would be interested in mouse and human targets for flow cytometry and IHC. Is there anything else, I should be aware of? Problems, tips, etc.?
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