faq

Q:	I want to stimulate human primary T-cells with anti-CD3 and anti-CD28. Do you have a detailed protocol for this?
A:	Yes, the detailed protocol is part of our 4D-Nucleofector® Protocol for stimulated Human T Cells. For optimal stimulation of the transfected cells we recommend coating 96-wells plates for stimulation [Nunc Immuno™ Plate C96 Maxi Sorp™, Cat. No. 430 341] with 50 µl of a solution of Anti-Human CD3 MAB at a final concentration of 1 µg/ml and Anti-Human CD28 MAB at a final concentration of 2 µg/ml (or with a solution of a control antibody (purified mIgG(K)) at a final concentration 3 ug/ml). After incubation for 5 hours at 37°C/5% CO2 wells are washed three times with PBS containing 0.5% (w/v) BSA. Antibodies should be diluted in carbonate buffer (32 mM Na2CO3/16 mM NaHCO3) from 100 ng/µl stock solutions directly before use.

I want to stimulate human primary T-cells with anti-CD3 and anti-CD28. Do you have a detailed protocol for this?

Yes, the detailed protocol is part of our 4D-Nucleofector® Protocol for stimulated Human T Cells.

For optimal stimulation of the transfected cells we recommend coating 96-wells plates for stimulation [Nunc Immuno™ Plate C96 Maxi Sorp™, Cat. No. 430 341] with 50 µl of a solution of Anti-Human CD3 MAB at a final concentration of 1 µg/ml and Anti-Human CD28 MAB at a final concentration of 2 µg/ml (or with a solution of a control antibody (purified mIgG(K)) at a final concentration 3 ug/ml).

After incubation for 5 hours at 37°C/5% CO2 wells are washed three times with PBS containing 0.5% (w/v) BSA. Antibodies should be diluted in carbonate buffer (32 mM Na2CO3/16 mM NaHCO3) from 100 ng/µl stock solutions directly before use.

Related Cells:

T cell, human peripheral blood unstim.

Categories:

Primary Cells and Media

Transfection

Research Areas:

Cancer Research/Cell Biology

Immunotherapy / Hematology

Basic Research