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Characterization 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™ (Cat. No. 554724). The cells were fixed, permeabilized and subsequently stained with Alexa 488 conjugated anti-mouse IL-17A (clone TC11-18H10, Cat. No. 560220/560221) (middle and right panels) or Alexa 488 rat IgG1 isotype control (clone R3-34, Cat. no. 557720) (left panel) using Pharmingen's I/C staining protocol. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the unstimulated stained (right panel) and non-specific antibody (left panel) controls.
Characterization 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™ (Cat. No. 554724). The cells were fixed, permeabilized and subsequently stained with Alexa 488 conjugated anti-mouse IL-17A (clone TC11-18H10, Cat. No. 560220/560221) (middle and right panels) or Alexa 488 rat IgG1 isotype control (clone R3-34, Cat. no. 557720) (left panel) using Pharmingen's I/C staining protocol. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the unstimulated stained (right panel) and non-specific antibody (left panel) controls.
Characterization 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™ (Cat. No. 554724). The cells were fixed, permeabilized and subsequently stained with Alexa 488 conjugated anti-mouse IL-17A (clone TC11-18H10, Cat. No. 560220/560221) (middle and right panels) or Alexa 488 rat IgG1 isotype control (clone R3-34, Cat. no. 557720) (left panel) using Pharmingen's I/C staining protocol. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the unstimulated stained (right panel) and non-specific antibody (left panel) controls.
Characterization 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™ (Cat. No. 554724). The cells were fixed, permeabilized and subsequently stained with Alexa 488 conjugated anti-mouse IL-17A (clone TC11-18H10, Cat. No. 560220/560221) (middle and right panels) or Alexa 488 rat IgG1 isotype control (clone R3-34, Cat. no. 557720) (left panel) using Pharmingen's I/C staining protocol. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the unstimulated stained (right panel) and non-specific antibody (left panel) controls.
Characterization 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™ (Cat. No. 554724). The cells were fixed, permeabilized and subsequently stained with Alexa 488 conjugated anti-mouse IL-17A (clone TC11-18H10, Cat. No. 560220/560221) (middle and right panels) or Alexa 488 rat IgG1 isotype control (clone R3-34, Cat. no. 557720) (left panel) using Pharmingen's I/C staining protocol. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the unstimulated stained (right panel) and non-specific antibody (left panel) controls.
Characterization 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™ (Cat. No. 554724). The cells were fixed, permeabilized and subsequently stained with Alexa 488 conjugated anti-mouse IL-17A (clone TC11-18H10, Cat. No. 560220/560221) (middle and right panels) or Alexa 488 rat IgG1 isotype control (clone R3-34, Cat. no. 557720) (left panel) using Pharmingen's I/C staining protocol. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the unstimulated stained (right panel) and non-specific antibody (left panel) controls.
Characterization 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™ (Cat. No. 554724). The cells were fixed, permeabilized and subsequently stained with Alexa 488 conjugated anti-mouse IL-17A (clone TC11-18H10, Cat. No. 560220/560221) (middle and right panels) or Alexa 488 rat IgG1 isotype control (clone R3-34, Cat. no. 557720) (left panel) using Pharmingen's I/C staining protocol. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the unstimulated stained (right panel) and non-specific antibody (left panel) controls.
Characterization 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™ (Cat. No. 554724). The cells were fixed, permeabilized and subsequently stained with Alexa 488 conjugated anti-mouse IL-17A (clone TC11-18H10, Cat. No. 560220/560221) (middle and right panels) or Alexa 488 rat IgG1 isotype control (clone R3-34, Cat. no. 557720) (left panel) using Pharmingen's I/C staining protocol. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the unstimulated stained (right panel) and non-specific antibody (left panel) controls.
Characterization 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™ (Cat. No. 554724). The cells were fixed, permeabilized and subsequently stained with Alexa 488 conjugated anti-mouse IL-17A (clone TC11-18H10, Cat. No. 560220/560221) (middle and right panels) or Alexa 488 rat IgG1 isotype control (clone R3-34, Cat. no. 557720) (left panel) using Pharmingen's I/C staining protocol. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the unstimulated stained (right panel) and non-specific antibody (left panel) controls.
Characterization 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™ (Cat. No. 554724). The cells were fixed, permeabilized and subsequently stained with Alexa 488 conjugated anti-mouse IL-17A (clone TC11-18H10, Cat. No. 560220/560221) (middle and right panels) or Alexa 488 rat IgG1 isotype control (clone R3-34, Cat. no. 557720) (left panel) using Pharmingen's I/C staining protocol. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the unstimulated stained (right panel) and non-specific antibody (left panel) controls.
Characterization 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™ (Cat. No. 554724). The cells were fixed, permeabilized and subsequently stained with Alexa 488 conjugated anti-mouse IL-17A (clone TC11-18H10, Cat. No. 560220/560221) (middle and right panels) or Alexa 488 rat IgG1 isotype control (clone R3-34, Cat. no. 557720) (left panel) using Pharmingen's I/C staining protocol. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the unstimulated stained (right panel) and non-specific antibody (left panel) controls.
Characterization 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™ (Cat. No. 554724). The cells were fixed, permeabilized and subsequently stained with Alexa 488 conjugated anti-mouse IL-17A (clone TC11-18H10, Cat. No. 560220/560221) (middle and right panels) or Alexa 488 rat IgG1 isotype control (clone R3-34, Cat. no. 557720) (left panel) using Pharmingen's I/C staining protocol. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the unstimulated stained (right panel) and non-specific antibody (left panel) controls.
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For flow cytometry of cell suspensions from peripheral lymphoid tissues, it is recommended that the cells be pre-incubated with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No.553141/553142).
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.
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