CD154 (CD40 ligand) is also known as CD40L, gp39, TRAP and T-BAM. CD40 ligand is a 32-39 kD type II transmembrane glycoprotein. It is a member of the TNF superfamily and is expressed on activated T cells. It has been reported to be important for B cell costimulation following binding of its receptor, CD40. Additionally, binding of CD40L to CD40 on B cells promotes the secretion of immunoglobulin and Ig isotype switching. CD40L is also involved in the regulation of cytokine production by T cells.
CD40LG
Reactivity: Human
FACS, ELISA
Host: Mouse
Monoclonal
8H10F5
unconjugated
Application Notes
Optimal working dilution should be determined by the investigator.
Assay Procedure
Take 100 μL peripheral blood anticoagulated by EDTA and add to the bottom of 5 mLtube,
Add 10 μL labeled antibody to the bottom of flow tube mixing with the whole blood, incubate for 20 minutes at room temperature away from light,
Add 2 ml1×RBC lysis buffer, incubate for 10 minutes away from light after mixing, dissolve red blood cells (recommended: RBC lysing Solution 10×,Cat.: FXP001),
Sample tube is set to 1000 rpm centrifugation for 5 minutes, discard the supernatant,
Add 2 mLPBS wash buffer to resuspend the cells, then1000 rpm centrifugation for 5 minutes, discard the supernatant,
Add 0.5 mLPBS wash buffer to resuspend the cells and detect by flow cytometry (sample should be determined on the day on the machine and can also be added fixation overnight at 4 °C then measured).