PE Rat Anti-Mouse IL-12 (p40/p70)
Clone C15.6 (RUO)
- Brand BD Pharmingen™
- Concentration 0.2 mg/ml
- Isotype Rat IgG1
- Reactivity Mouse (QC Testing)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen CHO-expressed recombinant mouse IL-12 p70 protein
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The C15.6 monoclonal antibody specifically binds to both free and complexed (homodimer p80 and heterodimer p70) forms of the p40 subunit of mouse interleukin-12 (IL-12). The immunogen used to generate the C15.6 hybridoma was recombinant mouse IL-12 p70 protein. p40 has also been described as a subunit of IL-23 and thus it is possible that the C15.6 antibody will crossreact with IL-23.
- Format PE
- Excitation Source Blue 488 nm,Green 532 nm,Yellow/Green 561 nm
- Excitation Max 496 nm
- Emission Max 578 nm
R-phycoerythrin (PE) is an accessory photosynthetic pigment found in red algae. It exists in vitro as a 240-kDa protein with 23 phycoerythrobilin chromophores per molecule. This makes PE one of the brightest fluorochromes for flow cytometry applications, but its photobleaching properties make it unsuitable for fluorescence microscopy.
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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.
- 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.
Immunofluorescent Staining and Flow Cytometric Analysis: The C15.6 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-12 producing cells within mixed cell populations. The PE- and APC-conjugated C15.6 antibodies are especially suitable for these experiments (see figure). For optimal immunofluorescent staining and flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.5 µg mAb/million cells). For specific methodology, see The Immune Function Handbook at our website at www.bdbiosciences.com.
A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the PE-C15.6 antibody with excess ligand prior to staining, or 2) pre-block paraformaldehyde-fixed/saponin-permeabilized cells with unlabeled C15.6 antibody (e.g., Cat. No. 554477) prior to staining. The intracellular staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable rat IgG1 isotype control immunoglobulin for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse or human cells is PE-R3-34 immunoglobulin (Cat. No. 554685); use at comparable concentrations to antibody of interest.
ELISA: The purified C15.6 antibody (Cat. No. 551219) is useful as a capture antibody for a sandwich ELISA for measuring mouse IL-12 p40 protein levels. The purified C15.6 antibody can be paired with the biotinylated C17.8 antibody (Cat. No. 554476) as the detection antibody, with recombinant mouse IL-12 p40 protein (Cat. No. 554594) as the standard. For testing mouse IL-12 p40 in complex biological fluids such as serum or plasma, our mouse IL-12 specific OptEIA™ sandwich ELISA set is recommended (Cat. No. 555165).