BD Pharmingen™ Alexa Fluor® 700 Rat Anti-Mouse IL-17A
Clone TC11-18H10 (RUO)
Flow cytometric analysis of IL-17A-producing cells within a stimulated mouse EL4 thymoma cell population. EL4 cells were stimulated (left and middle panels) or unstimulated (right panel) with PMA (50 ng/ml final concentration; Sigma, Cat. No. P-8139) and Ionomycin (1000 ng/ml final concentration; Sigma, Cat. No. I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724) for 5 hours. The cells were fixed, permeabilized and subsequently stained with Alexa Fluor® 700 Rat Anti-Mouse IL-17A (Cat. No. 560820; middle and right panels) or Alexa Fluor® 700 Rat IgG1 κ Isotype Control (Cat. No. 558001; left panel) using BD Biosciences' protocol for Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis. Bivariate flow cytometric dot plots showing correlated expression patterns of IL-17A (Alexa Fluor® 700) versus cellular autofluorescence (FL1) were derived from gated events with the forward and side light-scatter characteristics of viable cells. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control (FL1) and the Alexa Fluor® 700 Rat IgG1 κ Isotype Control staining. Flow cytometry was performed on a BD™ LSR II Flow Cytometer System.
.png){kind=spacer}
Flow cytometric analysis of IL-17A-producing cells within a stimulated mouse EL4 thymoma cell population. EL4 cells were stimulated (left and middle panels) or unstimulated (right panel) with PMA (50 ng/ml final concentration; Sigma, Cat. No. P-8139) and Ionomycin (1000 ng/ml final concentration; Sigma, Cat. No. I-0634) in the presence of BD GolgiStop™ Protein Transport Inhibitor (Cat. No. 554724) for 5 hours. The cells were fixed, permeabilized and subsequently stained with Alexa Fluor® 700 Rat Anti-Mouse IL-17A (Cat. No. 560820; middle and right panels) or Alexa Fluor® 700 Rat IgG1 κ Isotype Control (Cat. No. 558001; left panel) using BD Biosciences' protocol for Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis. Bivariate flow cytometric dot plots showing correlated expression patterns of IL-17A (Alexa Fluor® 700) versus cellular autofluorescence (FL1) were derived from gated events with the forward and side light-scatter characteristics of viable cells. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control (FL1) and the Alexa Fluor® 700 Rat IgG1 κ Isotype Control staining. Flow cytometry was performed on a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
IL-17; Il17; IL-17A; Il17a; CTLA-8; Ctla8; Interleukin-17A; interleukin 17A
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG1, κ
Recombinant Mouse IL-17A Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2034015
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 700 under optimum conditions, and unreacted Alexa Fluor® 700 was removed.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Alexa Fluor® 700 has an adsorption maximum of ~700nm and a peak fluorescence emission of ~720nm. Before staining cells with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- 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.
- 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/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Fixation and Permeabilization Solution RUO
Size
125 mL
Cat No.
554722
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Protein Transport Inhibitor (Containing Monensin) RUO
Size
0.7 mL
Cat No.
554724
Alexa Fluor® 700 Rat IgG1 κ Isotype Control RUO
Size
0.1 mg
Cat No.
558001
560820 Rev. 1
Antibody Details
TC11-18H10
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.
560820 Rev. 1
Format Details
Alexa Fluor™ 700
Alexa Fluor™ 700 dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 697 nm and an emission maximum (Em Max) at 719-nm. Alexa Fluor™ 700 is designed to be excited by the Red (627–640-nm) laser and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 700
Red 627-640 nm
697 nm
719 nm
560820 Rev.1
Citations & References
View product citations for antibody "560820" on CiteAb
Development References (5)
-
Dong C. Th17 cells: Current understanding of their generation and regulation. Eur J Immunol. 2009; 39(3):640-644. (Biology). View Reference
-
Kennedy J, Rossi DL, Zurawski SM, et al. Mouse IL-17: a cytokine preferentially expressed by alpha beta TCR + CD4-CD8-T cells. J Interferon Cytokine Res. 1996; 16(8):611-617. (Biology). View Reference
-
Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology). View Reference
-
Schwarzenberger P, La Russa V, Miller A, et al. IL-17 stimulates granulopoiesis in mice: use of an alternate, novel gene therapy-derived method for in vivo evaluation of cytokines. J Immunol. 1998; 161(11):6383-6389. (Biology). View Reference
-
Yen D, Cheung J, Scheerens H et al. IL-23 is essential for T cell–mediated colitis and promotes inflammation via IL-17 and IL-6. J Clin Invest. 2006; 116(5):1310-1316. (Biology). View Reference
560820 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.