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PE Rat Anti-Human IL-12 p70
PE Rat Anti-Human IL-12 p70

Specific control for intracellular staining of human IL-12 p70-producing cells. Pharmingen's HiCK-3 positive control cells (Cat. No. 555063) were intracellulary stained using Pharmingen's intracellular staining protocol.  The panels show intracellular staining with PE-conjugated antihuman IL-12 p70 (0.06 µg) and the pre-blocking of that staining with purified anti-human IL-12 p70 (5.0 µg, see right panel).

Specific control for intracellular staining of human IL-12 p70-producing cells. Pharmingen's HiCK-3 positive control cells (Cat. No. 555063) were intracellulary stained using Pharmingen's intracellular staining protocol.  The panels show intracellular staining with PE-conjugated antihuman IL-12 p70 (0.06 µg) and the pre-blocking of that staining with purified anti-human IL-12 p70 (5.0 µg, see right panel).

Product Details
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BD Pharmingen™
Human (QC Testing)
Rat IgG1, κ
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_397222
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


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

Immunofluorescent Staining and Flow Cytometric Analysis: The 20C2 antibody is useful for immunofluorescent staining and flow cytometric analysis of mixed cell populations to identify and enumerate IL-12 p70 producing cells. This 100 Test Size formulation of the PE-conjugated 20C2 antibody has been pre-titrated to assure effective intracellular detection of human IL-12 p70 using 20 µl/1 x 10^6 cells.

A suitable rat IgG1 isotype control for assessing the level of fluorochrome associate background staining on paraformaldehyde-fixed/saponin-permeabilized human cells is also available in a 100 Test Size formulation PE-R3-34 immunoglobulin (Cat. No. 559318).  A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the fluorochrome-conjugated 20C2 antibody with excess ligand (e.g., recombinant human IL-12 p70, Cat. No. 554613) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabeled 20C2 antibody (Cat. No. 557018) prior to staining. The intracellular staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe.

Important Note: This pre-titered antibody solution does not contain a cell permeabilization agent. It is necessary to include a cell permeabilization agent when using the pre-titered antibody solution to stain fixed and permeabilized cells. Perm/Wash™ Buffer (Cat. No. 554723) contains the permeabilization agent saponin and is useful for this purpose.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. 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.
  6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
559325 Rev. 2
Antibody Details
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20C2

The 20C2 antibody reacts with the biologically active form of human IL-12 p70 heterodimer, but not with the p40 monomer or homodimer.

559325 Rev. 2
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
559325 Rev.2
Citations & References
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Development References (2)

  1. Gately MK, Chizzonite R, Presky DH. Measurement of Human and Mouse Interleukin-12. In: Cooligan J, Kruisbeek A, Margulies D, Shevach E, Storber W, ed. Current Protocols in Immunology. New York: John Wiley and Sons; 1995:6-16.
  2. Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology). View Reference
559325 Rev. 2

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.