BD, 552536, BD Pharmingen™ Biotin Mouse Anti-Human CD120a

Alternative Name: TNF Receptor Type I; TNFR1; TNFR55; TNFR60; TNFRSF1A
Reactivity: Human (QC Testing)
Isotype: Mouse BALB/c IgG2a, κ
Immunogen: Human CD120a Recombinant Protein
Application: Flow cytometry (Routinely Tested), ELISA Detection (Tested During Development)
Concentration: 0.5 mg/ml
RRID: AB_393945
Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.
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. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.
Recommended Assay Procedures: Recommended Assay Procedures Immunofluorescent Staining and Flow Cytometric Analysis: Biotin Mouse Anti-Human CD120a (Cat. No. 550900) can be used for immunofluorescent staining (≤ 1 µg antibody/10×6 cells) and flow cytometric analysis of nucleated human cells to measure their expressed levels of surface TNFRI. A two-layer staining protocol is recommended for maximizing the detection of TNFRI expressed by cells as detailed in the figure legend. As a demonstration of specificity, the binding of MABTNFR1-B1 to TNFRI is inhibited when human TNFRI+ target cells are preincubated with saturating levels of recombinant human TNF at 4°C, i.e., when the TNFRI are bound with ligand. Based on our data, a recombinant human TNF level above 50 ng/10×6 cells is sufficient to completely inhibit the binding of MABTNFR1-B1 (0.06 µg/10×6 cells). Please note also that as a consequence of in vivo or in vitro activation, cell surface TNFRI can either be shed by cells or transiently expressed at higher levels. As a result, cellular activation can affect the cell's overall expressed level of surface TNFRI. ELISA Detection : Biotin Mouse Anti-Human CD120a serves as the detection antibody in a sandwich ELISA for measuring human soluble TNFRI protein levels. Biotin Mouse Anti-Human CD120a antibody can be paired with the Purified Mouse Anti-Human CD120a (Cat No. 552535) using recombinant human soluble TNFRI as the standard. This detection antibody should be titrated between 0.5 - 1 µg/ml to determine its optimal concentration for ELISA detection. To obtain linear standard curves, doubling dilutions of recombinant soluble human TNFRI ranging from 200 to 15 pg/ml are recommended for inclusion in each ELISA plate.  For specific methodology, please visit the protocols section under "ELISA and ELISPOT" at our website, http://www.bdbiosciences.com/us/s/resources.