BD, 554401, BD Pharmingen™ PE Rat Anti-Mouse IL-6

Reactivity: Mouse (QC Testing)
Isotype: Rat IgG1
Immunogen: Mouse IL-6 Recombinant Protein
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 0.2 mg/ml
RRID: AB_395367
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 PE-conjugated MP5-20F3 antibody (Cat. No. 554401) can be used for multicolor immunofluorescent staining and flow cytometric analyses to identify and enumerate IL-6-producing cells within mixed cell populations (see images). For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.25 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 conjugated MP5-20F3 antibody with ligand (e.g., recombinant mouse IL-6; Cat No. 554582) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabeled MP5-20F3 antibody (Cat. No. 554400) prior to staining. The 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 mouse or human cells is purified R3-34 (Cat. No. 554685); use at comparable concentrations to antibody of interest (e.g., ≤ 0.25 µg mAb/1 million cells). Other applications for this clone in additional formats available are: Neutralization : The NA/LE™ MP5-20F3 antibody (Cat. No. 554398) is useful for neutralization of mouse IL-6 bioactivity. ELISA Capture : The purified MP5-20F3 antibody (Cat. No. 554401) is useful as a capture antibody for a sandwich ELISA for measuring mouse IL-6 protein levels with the biotinylated detection antibody (Cat. No. 554402) and recombinant mouse IL-6 (Cat. No. 554582) as a standard. Western Blot : The MP5-20F3 antibody has been reported to be useful for Western blotting. Please note that this application is not 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.