Interested in promotions? | Click here >>
39901
IL-17A (D1X7L) Rabbit mAb (Mouse Specific) (PE Conjugate)
Antibody Conjugates
Monoclonal Antibody

IL-17A (D1X7L) Rabbit mAb (Mouse Specific) (PE Conjugate) #39901

Reviews ()
Citations (2)
Filter:
  1. F
Flow Cytometry Image 1: IL-17A (D1X7L) Rabbit mAb (Mouse Specific) (PE Conjugate)

Flow cytometric analysis of EL4 cells, untreated (left) or treated with TPA #4174 (50 ng/ml), Ionomycin, Calcium Salt #9995 (500 ng/ml), and Brefeldin A #9972 (500 ng/ml, overnight; right), using IL-17A (D1X7L) Rabbit mAb (Mouse Specific) (PE Conjugate).

To Purchase # 39901S
Product # Size Price
39901S
100 µl  (50 tests) $ 314

Supporting Data

REACTIVITY M
SENSITIVITY Endogenous
MW (kDa)
Source/Isotype Rabbit IgG

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Description

This Cell Signaling Technology antibody is conjugated to phycoerythrin (PE) and tested in-house for direct flow cytometry analysis in mouse cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated IL-17A (D1X7L) Rabbit mAb (Mouse Specific) #13838.

Product Usage Information

Application Dilution
Flow Cytometry 1:50

Storage

Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

Protocol

PRINT

View >Collapse >

Flow Cytometry General Protocol

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 20X Phosphate Buffered Saline (PBS): (#9808) To prepare 1 L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix.
  2. 16% Formaldehyde (methanol free).
  3. 0.1% Saponin in 1X PBS.
  4. Incubation Buffer: Dissolve 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.
  5. Recommended Anti-Rabbit secondary antibodies:

B. Fixation

  1. Collect cells by centrifugation and aspirate supernatant.
  2. Resuspend cells in 0.5–1 ml 1X PBS. Add appropriate volume of 16% Formaldehyde to obtain a final concentration of 4% (e.g. 750 μl PBS + 250 μl 16% Formaldehyde).
  3. Fix for 10 min in a 37°C water bath.
  4. Chill samples on ice for 1 min.
  5. For extracellular staining with antibodies that do not require permeabilization, proceed to immunostaining (Section D) or store cells in 1X PBS with 0.1% sodium azide at 4°C; for intracellular staining, proceed to permeabilization (Section C).

C. Permeabilization

  1. Wash cells by centrifugation and resuspend in 1 ml 0.1% Saponin/1X PBS.
  2. Incubate 30 min at room temperature.
  3. Add 2 ml of 0.1% Saponin/1X PBS and wash by centrifugation.

D. Immunostaining

  1. Resuspend cells in 100 µl of primary antibody diluted in 0.1% Saponin/1X PBS. See individual antibody datasheet or product webpage for the appropriate antibody dilution.
  2. Incubate for 1 hr at room temperature.
  3. Wash by centrifugation in 2 ml 0.1% Saponin/1X PBS.
  4. Resuspend cells in fluorochrome-conjugated secondary antibody or fluorochrome-conjugated avidin, diluted in 0.1% Saponin/1X PBS at the recommended dilution.
  5. Incubate for 30 min at room temperature.
  6. Wash by centrifugation in 2 ml 0.1% Saponin/1X PBS.
  7. Resuspend cells in 0.5 ml 1X PBS and analyze on flow cytometer; alternatively, for DNA staining, proceed to optional DNA dye (Section E).

E. Optional DNA Dye

  1. Resuspend cells in 0.5 ml of DNA dye (e.g. Propidium Iodide (PI)/RNase Staining Solution #4087).
  2. Incubate for at least 30 min at room temperature.
  3. Analyze cells in DNA staining solution on flow cytometer.

posted June 2014

Protocol Id: 604

Specificity / Sensitivity

IL-17A (D1X7L) Rabbit mAb (Mouse Specific) (PE Conjugate) recognizes endogenous levels of total mouse IL-17A protein.

Species Reactivity:

Mouse

Species predicted to react based on 100% sequence homology:

Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val49 of mouse IL-17A protein.

Background

The IL-17 family of cytokines consists of IL-17A-F, and their receptors include IL-17RA-RE (1). IL-17 cytokines are produced by a variety of cell types including the Th17 subset of CD4+ T cells, as well as subsets of γδ T cells, NK cells, and NKT cells (2). IL-17A and IL-17F, the most well-studied of the IL-17 cytokines, contribute to fungal and bacterial immunity by inducing expression of proinflammatory cytokines, chemokines, and antimicrobial peptides (2). In addition, IL-17A contributes to the pathogenesis of several autoimmune diseases (3). IL-17E promotes Th2 cell responses (4). The roles of IL-17B, IL-17C, and IL-17D are less clear, however these family members also appear to have the capacity to induce proinflammatory cytokines (1,5,6). IL-17 receptors have an extracellular domain, a transmembrane domain, and a SEFIR domain. They are believed to signal as homodimers, heterodimers, or multimers through their SEFIR domain by recruiting the SEFIR domain-containing adaptor Act1 (7). Unlike most cytokines that signal through Jak/STAT pathways, IL-17 signaling results in NF-κB activation (8).

  1. Gaffen, S.L. (2009) Nat Rev Immunol 9, 556-67.
  2. Iwakura, Y. et al. (2011) Immunity 34, 149-62.
  3. Hu, Y. et al. (2011) Ann N Y Acad Sci 1217, 60-76.
  4. Fort, M.M. et al. (2001) Immunity 15, 985-95.
  5. Yamaguchi, Y. et al. (2007) J Immunol 179, 7128-36.
  6. Li, H. et al. (2000) Proc Natl Acad Sci U S A 97, 773-8.
  7. Chang, S.H. et al. (2006) J Biol Chem 281, 35603-7.
  8. Shalom-Barak, T. et al. (1998) J Biol Chem 273, 27467-73.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.