IL-17A antibody

• Catalog Number: orb345240
• Category: Antibodies
• Description: IL-17A antibody
• Species/Host: Rat
• Clonality: Monoclonal
• Clone Number: 20B4.G10.F5
• Tested applications: ELISA, FC, Multiplex Assay, WB
• Reactivity: Mouse
• Isotype: IgG2a
• Immunogen: This Protein A purified monoclonal antibody was produced in rats by repeated immunizations with full length recombinant mouse IL-17A protein (produced in E.coli) followed by hybridoma development.
• Concentration: 2.94 mg/mL
• Dilution range: ELISA: 1:10,000, FC: 1:500, WB: 1:1000
• Form/Appearance: Liquid (sterile filtered)
• Purity: Anti-IL-17A is purified by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above. This antibody is specific for mouse Il-17a protein. Cross-reactivity with Il-17a from other sources has not been determined.
• Conjugation: Unconjugated
• UniProt ID: Q62386
• NCBI: NP_034682.1
• Storage: Store vial at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 µL). To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: rat anti-IL-17A antibody, rat anti-interleukin17A
• Note: For research use only
• Application notes: IL-17A monoclonal antibody has been tested for use in western blotting, flow cytometry and ELISA. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 23 kDa in size corresponding to the mature mouse Il17A protein, a non-glycosylated polypeptide chain consisting of 207 amino acids, by western blotting in appropriate cell lysate or extract.
• Expiration Date: 12 months from date of receipt.

Biorbyt monoclonal anti-IL-17A was used to detect IL-17A and separate Mouse CD4+ Cells by flow cytometry. Mouse CD4+ T cells were isolated from freshly dissected spleen by centrifugation in T cell separation media and selected by magnetic separation. Cells were grown on plates coated with anti-CD3 antibody, and stimulated with: 5 µg/mL anti-CD28, 10 ng/mL IL-1beta, 50 ng/mL mouse IL-6, 1 ng/mL TGFbeta1 and 10 µg/mL anti-mouse IFN gamma over 8-10 days of culture. Cells were incubated for 15-20 minutes with addition of rat anti-mouse CD4 APC at a concentration of 0.125 µg/mL, washed, fixed and permeabilized and incubated with Biorbyt Rat anti-mouse IL-17A monoclonal Antibody (orb345239) or controls as shown. Cells were washed, incubated in streptavidin conjugated PE, fixed and analyzed by Flow cytometry. Shown here are results for Biorbyt's monoclonal anti mouse IL-17A antibody (orb345239).

Biorbyt monoclonal anti-IL17A was used to detect IL-17A and separate Mouse CD4+ Cells by flow cytometry. Mouse CD4+ T cells were isolated from freshly dissected spleen by centrifugation in T cell separation media and selected by magnetic separation. Cells were grown on plates coated with anti-CD3 antibody, and stimulated with: 5 µg/mL anti-CD28, 10 ng/mL IL-1beta, 50 ng/mL mouse IL-6, 1 ng/mL TGFbeta1 and 10 µg/mL anti-mouse IFN gamma over 8-10 days of culture. Cells were incubated for 15-20 minutes with addition of rat anti-mouse CD4 APC at a concentration of 0.125 µg/mL, washed, fixed and permeabilized and incubated with Biorbyt Rat anti-mouse IL-17A monoclonal Antibody (orb345239) or controls as shown. Cells were washed, incubated in streptavidin conjugated PE, fixed and analyzed by Flow cytometry. Shown here are results for positive and negative controls.

Western Blot showing detection of Mouse IL-17A. 100 ng of Mouse IL-17A was run on a 4-20% gel and transferred to 0.45 µM nitrocellulose. After blocking with 1% BSA-TTBS (p/n orb348540, diluted to 1X) 30 min at 20°C, Anti-Mouse IL-17A (RAT) Antibody (p/n orb345239) was used at 1:1000 in 1% BSA-TTBS over night at 4°C. Peroxidase conjugated Rabbit Anti-mouse secondary antibody (p/n orb347544) was diluted in Blocking Buffer for Fluorescent Western Blotting (p/n orb348637) at 1:40000 for 30 min at 20°C. Band indicates correct 23 kDa molecular weight position expected for Mouse IL-17A.