BD, 554489, BD Pharmingen™ PE Rat Anti-Human IL-5

Reactivity: Human (QC Testing)
Isotype: Rat IgG2a
Immunogen: Recombinant human IL-5
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 0.2 mg/ml
RRID: AB_395427
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 JES1-39D10 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate human IL-5 producing cells within mixed cell populations. The PE-conjugated JES1-39D10 antibody (Cat. No. 554489) is especially suitable for these experiments (see Figure). 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 useful control for demonstrating specificity of staining is either of the following: 1) pre-block the PE-JES1-39D10 antibody with excess ligand (human IL-5 , Cat. No. 554606) prior to staining, or 2) pre-block paraformaldehyde-fixed/saponin-permeabilized cells with unlabeled JES1-39D10 antibody (Cat. No. 554488) prior to staining. The intracellular staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable rat IgG2a isotype control immunoglobulin for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse or human cells is the PE-R35-95 antibody (Cat. No. 554689); use at comparable concentrations to antibody of interest. ELISA Capture: The purified JES1-39D10 antibody (Cat. No. 554488) is useful as a capture antibody for a sandwich ELISA for measuring human IL-5 protein levels. WB: The purified JES1-39D10 antibody (Cat. No. 554487) has been found useful for Western blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen.
Product Notices: Product Notices Since applications vary, each investigator should titrate the reagent to obtain optimal results. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.