PE Rat Anti-Mouse IL-17A
Clone TC11-18H10 (RUO)
- Brand BD Pharmingen™
- Concentration 0.2 mg/ml
- Isotype Rat LEW, also known as Lewis IgG1, κ
- Reactivity Mouse (QC Testing)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Recombinant Mouse IL-17A Protein
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The TC11-18H10 monoclonal antibody specifically binds to recombinant and natural mouse IL-17A proteins. IL-17A, also known as CTLA-8, is a T cell-derived cytokine that promotes inflammatory responses. Mouse IL-17A is a proinflammatory cytokine that can induce the release of IL-6 by mouse stromal cells. It has been shown to support the growth of hemopoietic progenitors in vitro; it can also stimulate granulopoiesis in vivo. The TC11-18H10 antibody has been reported to neutralize IL-17A activity. Recent studies have shown that IL-17A is produced by a unique subset of Th17 cells that develop along a pathway distinct from the Th1- and Th2- cell differentiation pathways. The mouse IL-17A cDNA was isolated from a cDNA library generated from TCRαβ+CD4-CD8- thymocytes.
- Format PE
- Excitation Source Blue 488 nm,Green 532 nm,Yellow/Green 561 nm
- Excitation Max 496 nm
- Emission Max 578 nm
R-phycoerythrin (PE) is an accessory photosynthetic pigment found in red algae. It exists in vitro as a 240-kDa protein with 23 phycoerythrobilin chromophores per molecule. This makes PE one of the brightest fluorochromes for flow cytometry applications, but its photobleaching properties make it unsuitable for fluorescence microscopy.
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Preparation and Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
Recommended Assay Procedure:
Immunofluorescent Staining and Flow Cytometric Analysis: The TC11-18H10 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-17A producing cells within mixed cell populations. The PE-conjugated TC11-18H10 antibody (Cat. No. 559502) is especially suitable for these experiments (see image). For optimal immunofluorescent staining and flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.25 µg mAb/million cells). For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on intracellular staining in the Immune Function Handbook..
A useful control for demonstrating the specificity of staining is to pre-block paraformaldehyde-fixed/saponin-permeabilized target cells with unlabeled TC11-18H10 antibody (Cat. No. 559501) prior to staining. The intracellular staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable rat IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse is PE-R3-34 (Cat. No. 554685); use at comparable concentrations to antibody of interest (e.g., ≤ 0.25 µg mAb/1 million cells).
1. ELISA Capture: The purified TC11-18H10 antibody (Cat. No. 555068) is useful as a capture antibody for a sandwich ELISA for measuring mouse IL-17A protein levels. Purified TC11-18H10 antibody can be paired with the biotinylated TC11-8H4 (Cat. No.555067) antibody as the detecting antibody, with recombinant mouse IL-17A protein as the standard. The purified TC11-18H10 antibody should be titrated from 0.5 µg/ml to 2.0 µg/ml to determine its optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of mouse IL-17A ranging from ~2,000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on ELISA in the Immune Function Handbook.
2. Western blotting: The TC11-1810 antibody has been found useful for Western Blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen.
3. Neutralization: TCII-18H10 has been shown to be a neutralizing antibody. Please note that this application is not routinely tested at BD Biosciences.