BD, 554484, BD Pharmingen™ FITC Rat Anti-Human IL-4

Reactivity: Human (QC Testing)
Isotype: Rat IgG1
Immunogen: Purified Recombinant Human IL-4
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 0.5 mg/ml
RRID: AB_395423
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 FITC under optimum conditions, and unreacted FITC was 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 Cytometry Analysis: The FITC-conjugated MP4-25D2 antibody can be used for multicolor immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-4-producing cells within mixed cell populations (see image, left panel). For optimal immunofluorescent staining for flow cytometric analysis, this antibody should be titrated (≤ 0.5 µg mAb/million cells). A useful control for demonstrating specificity of staining is the following: pre-block the fixed/permeabilized cells with unlabelled MP4-25D2 antibody (Cat. No. 554482) prior to staining. The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable rat IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells is FITC-R3-34 immunoglobulin (Cat. No. 554684); use at comparable concentrations to antibody of interest (e.g. , ≤ 0.5 µg mAb/ 1 million cells). 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. ELISA Detection: The biotinylated MP4-25D2 (Cat. No. 554483) antibody is useful as a detection antibody for a sandwich ELISA that measures human IL-4 protein levels. Biotinylated MP4-25D2 antibody can be paired with the purified 8D4-8 antibody (Cat. No. 554515) as the capture antibody, with recombinant human IL-4 protein (Cat. No. 554605) as the standard. For assay of IL-4 in serum or plasma, the BD OptEIA™ Human IL-4 ELISA Set (Cat. No. 555194) or  BD OptEIA™ Human IL-4 ELISA Kit (Cat. No. 550614) is recommended. Neutralization: The NA/LE format of the MP4-25D2 antibody (Cat. No. 554481) has been reported to be useful for neutralization of human IL-4 bioactivity. A suitable NA/LE™ rat IgG1 isotype control to match the NA/LE™ MP4-25D2 antibody is the R3-34 antibody, Cat. No. 554683.