BD, 554544, BD Pharmingen™ FITC Rat Anti-Human IL-6

Reactivity: Human (QC Testing)
Isotype: Rat IgG1
Immunogen: Human IL-6 Recombinant Protein
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 0.5 mg/ml
RRID: AB_395468
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 Cytometric Analysis: The PE conjugated MQ2-13A5 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-6 producing cells within mixed cell populations (see image). For optimal immunofluorescent staining for flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.5 µg mAb/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. A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the FITC-MQ2-13A5 antibody with excess ligand (e.g., recombinant human IL-6; Cat No. 550071) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabelled MQ2-13A5 antibody (Cat. No. 554543) prior to staining. A suitable rat IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells is FITC-R3-34 (Cat. No. 554684); use at comparable concentrations to antibody of interest (e.g., ≤ 0.5 µg mAb/1 million cells). OTHER APPLICATIONS ELISA Capture: The purified MQ2-13A5 antibody (Cat. No. 554543) is useful as a capture antibody for a sandwich ELISA for measuring human IL-6 protein levels.  Purified MQ2-13A5 antibody can be paired with the biotinylated MQ2-39C3 antibody (Cat. No. 554546) as the detecting antibody, with recombinant human IL-6 protein (Cat. No. 550071) as the standard. For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on ELISA in the Immune Function Handbook. For testing IL-6 in serum or plasma, our OptEIA™ Human IL-6 ELISA Set (Cat.No. 550799) or OptEIA Human IL-6 ELISA Kit (Cat. No. 550799) is recommended. Neutralization: The no azide/low endotoxin formulation of the MQ2-13A5 antibody (Cat. No. 554541) is useful for neutralization of human IL-6 bioactivity. Western blot: The MQ2-13A5 antibody (Cat. No. 554543) has been found useful for Western blot. 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. 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.