IL-17A Monoclonal Antibody (eBio64DEC17), Brilliant Violet™ 786, eBioscience™, Invitrogen™

Description

• IL-17A Monoclonal antibody specifically detects IL-17A in Human samples
• It is validated for Flow Cytometry Description: The eBio64DEC17 antibody reacts with human IL-17A
• The eBio64DEC17 antibody is a neutralizing antibody
• Interleukin-17A (IL-17A) 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
• The cDNA encoding human IL-17A was isolated from a library of CD4+ T cells; the encoded protein exhibits 72 percent amino acid identity with HVS13 , an open reading frame from a T lymphotropic Herpesvirus saimiri, and 63 percent with mouse CTLA-8 (cytotoxic T-lymphocyte associated antigen-8)
• Human IL-17A exists as glycosylated 20-30 kD homodimers
• High levels of IL-17A homodimer are produced by activated peripheral blood CD4+ T-cells
• IL-17A enhances expression of the intracellular adhesion molecule-1 (ICAM-1) in human fibroblasts
• 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 IL-17A receptor
• 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 Reported: This eBio64DEC17 antibody has been reported for use in intracellular staining followed by flow cytometric analysis
• 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 <a href='https://www.thermofisher.com/flowprotocols'>Flow Protocols</a>
• This may be used at 5 μL (0.5 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
• Brilliant Violet™ 786 (BV786) is a tandem dye that emits at 786 nm and is intended for use on cytometers equipped with a violet (405 nm) laser
• Please make sure that your instrument is capable of detecting this fluorochrome
• 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-4401-42) 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 # 00-8222-49) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) 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 Violet™ 786 (BV786) is not compatible with methanol-based fixation
• Excitation: 407 nm; Emission: 786 nm; Laser: Violet Laser
• BRILLIANT VIOLET™ is a trademark or registered trademark of Becton, Dickinson and Company or its affiliates, and is used under license
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