IL-12/IL-23 p40 Monoclonal Antibody (C8.6), Brilliant Violet™ 421, eBioscience™

Product Specific Information

Description
The C8.6 antibody reacts with the p40 subunit of human IL-12 p70 and human IL-23. IL-12 is a 70 kDa heterodimeric cytokine composed of two covalently linked chains, p40 and p35, and secreted primarily by activated monocytes, macrophages, and dendritic cells. IL-23 is a biologically-related, heterodimeric cytokine composed of disulfide-linked p40 and p19 subunits.

Applications Tested
This C8.6 antibody has been pre-diluted and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Please refer to "Staining Intracellular Antigens for Flow Cytometry, Protocol A: Two step protocol for intracellular (cytoplasmic) proteins" located at Flow Protocols. This may be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Blocking Buffers
When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) or Brilliant Stain Buffer (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information.

Fixation
• Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation.
• Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 407 nm; Emission: 423 nm; Laser: Violet Laser.

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Target Information

IL-12 is an interleukin produced by dendritic cells, neutrophils, macrophages and human lymphoblastoid cells as a result of antigenic stimulation. IL-12 is a heterodimeric cytokine encoded by IL-12A (p35) and IL-12B (p40). IL-23 is a heterodimeric cytokine that shares a subunit with IL-12 (IL-12B) along with an IL-23A (IL-23p19) subunit. Functionally, IL-12 increases the expression of vascular endothelial adhesion molecule-1 on the endothelium, directs leukocytes towards tumors and facilitates the ischemic-hemorrhagic necrosis of the tumor tissue. In comparison, IL-23 plays an important role in stimulating memory T-cells, and is required for the induction, expansion, maintenance and downstream effector functions of Th17 cells, which play a vital role in upregulating neutrophil chemokines and various pro-inflammatory cytokines.