BD, 554387, BD Pharmingen™ Purified Rat Anti-Mouse IL-4

Reactivity: Mouse (QC Testing)
Isotype: Rat IgG2b
Immunogen: Recombinant Mouse IL-4
Application: ELISA Capture, Intracellular block/flow cytometry (Routinely Tested), Neutralization (Tested During Development), Western blot (Reported)
Concentration: 0.5 mg/ml
RRID: AB_398544
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. Store undiluted at 4°C.
Recommended Assay Procedures: Recommended Assay Procedures 1. Blocking Control for Intracellular Staining: The purified BVD4-1D11 antibody (Cat. No. 554386) can be used as a blocking control to demonstrate specificity of IL-4 staining by conjugated-BVD4-1D11 and antibody. To perform this control, the fixed/permeabilized cells (~ 1 million) can be incubated with 1-10 µg of unlabeled BVD4-1D11 antibody (Cat. No. 554386) for 20 minutes at 4°C, prior to staining with conjugated-BVD4-1D11 antibody (e.g., 0.1 -0.5 µg mAb/1 million cells). The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on intracellular staining in the Immune Function Handbook. 2. ELISA Capture: The purified BVD4-1D11 antibody (Cat. No. 554387) is useful as a capture antibody for a sandwich ELISA for measuring mouse IL-4 protein levels in tissue culture supernatants. This antibody can be paired with the biotinylated BVD6-24G2 antibody (Cat. No. 554390) and recombinant mouse IL-4 (Cat. No. 550067) as the standard. For detecting IL-4 in serum or plasma, the BD OptEIA™  Mouse IL-4 ELISA Set (Cat. No. 555232) is recommended. Note: Purified BVD4-1D11 antibody has been found to yield higher background in ELISA than the alternative mouse IL-4 ELISA capture antibody, clone 11B11 (Cat. No. 554434). To overcome this high background, the following procedures specific for clone BVD4-1D11 are recommended: 1) Titrate BVD4-1D11 capture antibody (1 -4 µg/ml) versus biotinylated BVD6-24G2 detection antibody (0.1 - 1.0 µg/ml; i.e. lower than usual detection antibody concentration). Separate blanks must be used for each concentration of biotinylated detection antibody used. 2) Use twice the number of recommended washes at all steps of ELISA protocol. 3. Neutralization: The NA/LE™ BVD4-1D11 antibody (Cat. No. 554385) is useful for neutralization of mouse IL-4 bioactivity. 4. WB: The BVD4-1D11 antibody has been reported to be useful for Western blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen.