TotalSeq™-B1150 anti-human IL-12/IL-23 p40 Antibody

Product Details

Verified Reactivity

Human

Antibody Type

Monoclonal

Host Species

Mouse

Immunogen

CHO-expressed, recombinant human IL-12 p70 heterod

Formulation

Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and EDTA

Preparation

The antibody was purified by chromatography and conjugated with TotalSeq™-B oligomer under optimal conditions.

Concentration

0.5 mg/mL

Storage & Handling

The antibody solution should be stored undiluted between 2°C and 8°C. Do not freeze.

Application

ICPG - Quality tested

Recommended Usage

Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis and the oligomer sequence is confirmed by sequencing. TotalSeq™-B antibodies are compatible with 10x Genomics Single Cell Gene Expression Solutions.

To maximize performance, it is strongly recommended that the reagent be titrated for each application, and that you centrifuge the antibody dilution before adding to the cells at 14,000xg at 2 - 8°C for 10 minutes. Carefully pipette out the liquid avoiding the bottom of the tube and add to the cell suspension. For Proteogenomics analysis, the suggested starting amount of this reagent for titration is ≤ 0.5 µg per million cells in 100 µL volume. Refer to the corresponding TotalSeq™ protocol for specific staining instructions.

Buyer is solely responsible for determining whether Buyer has all intellectual property rights that are necessary for Buyer's intended uses of the BioLegend TotalSeq™ products. For example, for any technology platform Buyer uses with TotalSeq™, it is Buyer's sole responsibility to determine whether it has all necessary third party intellectual property rights to use that platform and TotalSeq™ with that platform.

Application Notes

Flow Cytometry^3,5: The fluorochrome-labeled C11.5 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify IL-12 p40 (monomeric, heterodimeric, homodimeric) -producing cells within mixed cell populations. For intracellular cytokine staining protocol, please visit www.biolegend.com and click on the support section.
Neutralization⁶: The LEAF™ purified antibody (Endotoxin <0.1 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 501813).
Additional reported applications (for the relevant formats) include: Western blotting, ELISA Capture^1,2, ICFC Blocking, immunohistochemical staining⁴ of paraformaldehyde-fixed, saponin-treated tissue frozen sections, and immunocytochemistry.
Note: For testing human IL-12/IL-23 p40 (monomer, dimer) in serum or plasma, BioLegend's ELISA Max™ Sets (Cat. No. 430701 to 430706) are specially developed and recommended. For testing human IL-12 p70 in serum or plasma, BioLegend's ELISA Max™ Sets (Cat. No. 431701 to 431706) are specially developed and recommended.

Additional Product Notes

TotalSeq™ reagents are designed to profile protein levels at a single cell level following an optimized protocol similar to the CITE-seq workflow. A compatible single cell device (e.g. 10x Genomics Chromium System and Reagents) and sequencer (e.g. Illumina analyzers) are required. Please contact technical support for more information, or visit biolegend.com/totalseq.

The barcode flanking sequences are GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNNNNNNNNN (PCR handle), and NNNNNNNNNGCTTTAAGGCCGGTCCTAGC*A*A (capture sequence). N represents either randomly selected A, C, G, or T, and * indicates a phosphorothioated bond, to prevent nuclease degradation.

View more applications data for this product in our Scientific Poster Library.

Application References

(PubMed link indicates BioLegend citation)

1. D'Andrea A, et al. 1992. J. Exp. Med. 176:1387.
2. D'Andrea A, et al. 1993. J. Exp. Med. 178:1041.
3. Prussin C, et al. 1995. J. Immunol. Meth. 188:117.
4. Leifeld L, et al. 2002. Hepatology 36:1001.
5. Pekkari K, et al. 2001. Blood 97:3184.
6. Bolovan-Fritts CA, et al. 2004. J. Virol. 78:13173.

RRID

AB_2941588 (BioLegend Cat. No. 501825)

Antigen Details

Structure

Cytokine; monomer, heterodimer (p40:p35 or p40:p19) or homodimer (p40:p40)

Bioactivity

IL-12 p70 (p40:p35) induces IFN-?, TNF-a production in T and NK cells; costimulation of PBL proliferation; proliferation/differentiation of TH1 T lymphocytes. IL-23 (p40:p19) induces proliferation and production of IFN-? by human memory T cells, but not n

Cell Sources

Dendritic cells, monocytes/macrophages, B cells, T cells

Cell Targets

T cells, NK cells, PBL

Receptors

IL-12Rß1 binds p40; dimeric with IL-12Rß2 binds p35

Biology Area

Immunology, Innate Immunity

Molecular Family

Cytokines/Chemokines

Antigen References

1. Fitzgerald K, et al. Eds. 2001. The Cytokine FactsBook. Academic Press San Diego.
2. Quesniaux V. 1992. Research Immunol. 143:385.
3. Trinchieri G, et al. 1992 Prog. Growth Factor Res. 4:355.
4. Trinchieri G, et al. 1993 Immunol. Today. 14:335.
5. Oppmann B, et al. 2000 Immunity. 13:715.
6. Aggarwal S, et al. 2003 J. Biol. Chem. 278:1910.
7. Parham C, et al. 2002 J. Immunol. 168:5699.
8. Belladonna ML, et al. 2002 J. Immunol. 168:5448.
9. Lankford CS, et al. 2003 J. Leukoc. Biol. 73:49.

Regulation

Downregulated by IL-10; homodimeric p40 antagonistic to functional p70 heterodimer; p40 monomer has no function; p40 subunit in common with IL-23

Gene ID

3593 View all products for this Gene ID

UniProt

View information about IL-12/IL-23 p40 on UniProt.org

Related Protocols

• TotalSeq™-B or -C Cell Surface and Intracellular Cytoplasmic Protein Staining with 10x Chromium Single Cell Gene Expression Flex

Other Formats