BD, 559330, 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)
Vol. Per Test: 20 µl
RRID: AB_397227
Storage Buffer: Aqueous buffered solution containing BSA and ≤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. 559330) can be used for multicolor immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-10-producing cells within mixed cell populations (see image). This 100 Test Size formulation of the PE-conjugated JES3-19F1 antibody has been pre-titrated to assure effective intracellular detection of human IL-10 using 20 µl/1 x 10^6 cells. A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the conjugated JES3-19F1 antibody with ligand (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 also available in a 100 Test Size formulation PE-R35-95 (Cat. No. 559317). 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. 559330) 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).