BD, 553655, BD Pharmingen™ Purified NA/LE Hamster Anti-Mouse CD154

Alternative Name: CD40 Ligand; CD40L; gp39; Ly-62; Tnfsf5; T-BAM; HIGM1; IMD3
Reactivity: Mouse (QC Testing)
Isotype: Armenian Hamster IgG3, κ
Immunogen: Activated mouse Th1 clone D1.6
Application: Flow cytometry (Routinely Tested), Blocking, Immunohistochemistry (Reported)
Concentration: 1.0 mg/ml
RRID: AB_394975
Storage Buffer: No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2µm sterile filtered. Endotoxin level is ≤0.01 EU/µg (≤0.001 ng/µg) of protein as determined by the LAL assay.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. This preparation contains no preservatives, thus it should be handled under aseptic conditions.
Recommended Assay Procedures: Recommended Assay Procedures Flow cytometry: For the detection of mouse CD154 on activated peripheral T cells, it is strongly recommended that T cells be purified before activation. Mouse CD154 is transiently expressed on the surfaces of activated normal T cells and certain T cell clones with a maximal level detected 6-8 hours post-activation. Activation with immobilized anti-CD3e mAb ( e.g ., 145-2C11, Cat. No. 557306/553058, or 500A2, Cat. No. 553238) is sufficient to induce CD154 expression on CD4+ cells. It has been reported that CD8+ cells express CD154 only in response to PMA/Ionomycin treatment. Therefore, for detection of CD154, it is crucial to utilize the proper activation stimuli and to stain cells at the optimal time for CD154 expression. We recommend the use of biotinylated mouse anti-hamster IgG cocktail (Cat. No. 554010) followed by a "bright" second-step reagent, such as Streptavidin-PE (Cat. No. 554061), for optimal detection of CD154.
Product Notices: Product Notices Since applications vary, each investigator should titrate the reagent to obtain optimal results. Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.