BD, 554706, BD Pharmingen™ PE Rat Anti-Human IL-10

Alternative Name: Interleukin-10; CSIF; Cytokine synthesis inhibitory factor; TGIF
Reactivity: Human (QC Testing), Viral (Tested in Development)
Isotype: Rat IgG2a, κ
Immunogen: Recombinant Human IL-10
Application: Intracellular staining (flow cytometry) (Routinely Tested), ELISA Capture, Neutralization (Tested During Development)
Concentration: 0.2 mg/ml
RRID: AB_395521
Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. 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.
Recommended Assay Procedures: Recommended Assay Procedures 1. Immunofluorescent Staining and Flow Cytometric Analysis: The PE-conjugated JES3-19F1 antibody (Cat. No. 554706) can be used for multicolor immunofluorescent staining and flow cytometric analyses to identify and enumerate IL-10-producing cells within mixed cell populations (see image). For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.5 µg mAb/million cells). A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the conjugated JES3-19F1 antibody with ligand (e.g., recombinant human IL-10; Cat. No. 554611) prior to staining, or 2) pre-block the fixed/ permeabilized cells with unlabelled JES3-19F1 antibody (Cat. No. 554705) prior to staining. The staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe.  A suitable rat IgG2a isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells is PE-R35-95 (Cat. No. 554689); use at comparable concentrations to the antibody of interest (e.g., ≤ 0.5 µg mAb/1 million cells). BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application. 2. ELISA Capture : The purified JES3-19F1 (Cat. No. 554705) antibody is useful as a capture antibody for a sandwich ELISA for specifically measuring human IL-10 protein levels. Purified JES3-19F1 antibody can be paired with the biotinylated JES3-12G8 antibody (Cat. No. 554499) as the detection antibody and with recombinant human IL-10 protein (Cat. No. 554611) as the standard. For testing IL-10 in serum or plasma, our OptEIA™ set (Cat. No. 555134) is recommended. 3. Neutralization : The NA/LE™ format of clone JES3-19F1 (Cat. No. 554704) is useful for neutralization of human IL-10 bioactivity. A suitable NA/LE rat IgG2a isotype control to match the NA/LE JES3-19F1 antibody is the R35-95 antibody, (Cat. No. 554687).
Product Notices: Product Notices Since applications vary, each investigator should titrate the reagent to obtain optimal results. An isotype control should be used at the same concentration as the antibody of interest. 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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application. Please refer to http://regdocs.bd.com to access safety data sheets (SDS). Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.