IL-17A Monoclonal Antibody (eBio64DEC17), Brilliant Ultra Violet™ 737, eBioscience™

Product Specific Information

Description
The eBio64DEC17 antibody reacts with human IL-17A. The eBio64DEC17 antibody is a neutralizing antibody. IL-23-dependent, IL-17A-producing CD4+ T cells (Th-17 cells) have been identified as a unique subset of Th cells that develops along a pathway that is distinct from the Th1- and Th2- cell differentiation pathways. The hallmark effector molecules of Th1 and Th2 cells, e.g., IFN gamma and IL-4, have each been found to negatively regulate the generation of these Th-17 cells. Intracellular staining by eBio64DEC17 antibody identifies the same cell population as the eBio64CAP17 antibody, as can be seen in co-staining experiments using both antibodies.

Applications Tested
This eBio64DEC17 antibody has been pre-diluted and tested by intracellular staining followed by flow cytometric analysis of stimulated 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 (1.0 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.

Light sensitivity
This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

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.
• Our internal testing suggests that Brilliant Ultra Violet™ 737 (BUV737) is compatible with short-term methanol-based fixation, but should not be stored in buffers containing methanol for longer than one hour.

Excitation: 355 nm; Emission: 732 nm; Laser: Ultraviolet Laser.

BRILLIANT ULTRA VIOLET™ is a trademark or registered trademark of Becton, Dickinson and Company or its affiliates, and is used under license. Powered by Sirigen™.

Target Information

Interleukin-17A (IL-17A, CTLA-8) is a CD4+ T cell-derived cytokine that promotes inflammatory responses in cell lines and is elevated in rheumatoid arthritis, asthma, multiple sclerosis, psoriasis, and transplant rejection. IL-17A is a 32 kDa long, disulfide-linked homodimer consisting of 136 amino acids that is a member of a six-species family of proteins (IL-17A-17F) and signals through the IL-17 receptor (IL-17R/CDw217). High levels of IL-17A homodimer are produced by activated peripheral blood CD4+ T-cells, and IL-17A also enhances expression of the intracellular adhesion molecule-1 (ICAM-1) in human fibroblasts. In particular, human IL-17A also stimulates epithelial, endothelial, or fibroblastic cells to secrete IL-6, IL-8, G-CSF, and PGE2. In the presence of human IL-17A, fibroblasts can sustain the proliferation of CD34+ hematopoietic progenitors and induce maturation into neutrophils. Mouse, rat, and human IL-17A can induce IL-6 secretion in mouse stromal cells, indicating that all homologs can recognize the mouse receptor. IL-17A regulates the activities of NF-kappa B and mitogen-activated protein kinases, stimulates the expression of IL-6 and cyclooxygenase-2 (PTGS2/COX-2), and enhances the production of nitric oxide (NO).