CD 3 Antibody Protocols


The tissue expression of CD3 can be found in adaptive immune response like cancers and normal lymphoid tissues. It is used as a prostatic markers like endometrial, renal and breast cancers.

While running my previous company, I worked up a protocol for CD3 on Human and Canine tissues. The antibody from Novus Biologicals worked very well using human tonsil and canine lymph node.

This protocol is now available to purchase here.

Recently I saw a post on research gate asking about a protocol for CD3 on mouse tissues. The discussion stated that mouse tissue is difficult to produce results on and that no one had a protocol for mouse tissues. I thought, wow, I knew that was not the case because so many of our clients did stain mouse tissues for it. Even though my company did not have a CD3 protocol for mouse tissues, we sent projects out for the staining to a partner lab.

Since my company is no more, and I have the partner lab protocols, I will now share them here. The CD3 antibody is Abcam ab16667, a rabbit monoclonal (data sheet below). The following protocol is designed for paraffin embedded formalin fixed (FFPE) tissues but could easily be modified for frozen tissues. This has been used with ABC or HRP methods. Using sodium citrate pH 6.0 for 30 minutes with a a dilution of 1:150 for 60-90 minutes at 60C and a rabbit secondary antibody will produce professional results.

The above protocols have been written for hand staining. Additional information like deparaffinization, background staining, supporting reagents and secondary antibodies has been omitted due to personal preferences and or laboratory SOP’s. If you need further assistance with these steps, please ask.

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