Hi
I want to use pre-coated CD3 and CD28 mAb to activate T cells. I just realize there are different types and clones to choose. Based on your experience, which antibody works best？from which company and which clone？
Thanks a lot!
Hello Tao Zhou,
unfortunately you did not specify, if you wanted to stimulate murine or human T cells. For murine T cells you can use anti CD3 ab (clone 145 2C11) at a concentration of 5 µg/ml (plate bound) and anti CD28 ab (clone 37.51) at a concentration of 2 µg/ml (in solution). Always worked perfectly for me, irrespective if I looked at T cell proliferation, activation or cytokine production. For further details on distributors etc. please check
Article A role for the histone H2A deubiquitinase MYSM1 in maintenan...
I hope that helps!
All the best,
Michael
Hey,
I don't know if you want to stimulate murine or human T-Cells, but
we use OKT3 as anti-CD3 clone (I think the company is Bioxcell) for human T-Cells. It has a really good stimulatory capacity even if you use it soluble (in my hands half of the stimulatory capacitiy was reached around 500 pg/ml when using pbmcs as responder cells)
For anti-CD28, we use clone NA/LE from BD Pharmingen. (also soluble possible but i would recommend platebound)
Greetings
Coating should work fine for expansion of t cells and a strong stimulus.
We coat our Plates in PBS with 0,5 - 1 µg/ml anti-CD3 and 1-2 µg/ml anti CD28 (lowest concentrations work fine). However, I did'nt make any titrations yet.
Hello,
For human T cells, we use anti-CD3 precoated on plates (500 ng / 24 w plate well, IM1304, Beckman Coulter) and anti-CD28 in culture medium (500 ng/ml, IM1376,Beckman Coulter).
Br, Henna
for Human
try functional grade monoclonal from Miltenyi Biotec
CD 28 clone: 15E8
CD3 clone OKT3
http://www.miltenyibiotec.com/en/products-and-services/macs-flow-cytometry/reagents/antibodies-and-dyes/cd3-antibodies--functional-grade-human.aspx
http://www.miltenyibiotec.com/en/products-and-services/macs-flow-cytometry/reagents/antibodies-and-dyes/cd28-antibodies--functional-grade-human.aspx
or their activation kits easy to set
http://www.miltenyibiotec.com/en/products-and-services/macs-cell-culture-and-stimulation/stimulation-reagents/stimulation-based-on-macsibead-particles/t-cell-activation-expansion-kit-human.aspx
Robert Ose if i want to run the proliferated T cells to flow, will these cells still bind to a fluorescent CD3 antibody?
Daniel Xin Su , No, following stimulation, T cells downeregulte CD3.
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