BD, 554676, BD Pharmingen™ PE Rat Anti-Human IL-3

Reactivity: Human (QC Testing)
Isotype: Rat IgG1
Immunogen: Recombinant Human IL-3
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 0.2 mg/ml
RRID: AB_395504
Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures: Recommended Assay Procedures Immunofluorescent Staining and Flow Cytometric Analysis: The BVD3-1F9 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-3 producing cells within mixed cell populations. The PE-conjugated BVD3-1F9 antibody (Cat. No.  554676) is especially suitable for these studies.  For optimal immunofluorescent staining and flow cytometric analysis, this anti-cytokine antibody should be titrated ( ≤ 0.5 µg mAb/million cells). For specific methodology, please visit the protocols section or chapter on intracellular staining in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com. A suitable rat IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells is R3-34 (Cat. No. 554685); use at comparable concentrations to antibody of interest (e.g., ≤ 0.5 µg mAb/1 million cells). A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the conjugated BVD3-1F9 antibody with ligand (e.g., recombinant human IL-3; Cat No. 554604) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabeled BVD3-1F9 antibody prior to staining. The staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. ELISA : The biotinylated BVD3-1F9 antibody (Cat. No. 554674) is useful as a detection antibody for a sandwich ELISA for measuring human IL-3 protein levels.  Biotinylated BVD3-1F9 antibody can be paired with the purified BVD8-3G11 antibody (Cat. No. 554672) as the capture antibody, with recombinant human IL-3 protein (Cat. No. 554604) as the standard.