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BUV395 Rat Anti-Mouse IL-17A
BUV395 Rat Anti-Mouse IL-17A
Flow cytometric analysis of IL-17A-producing mouse EL-4 cells. Cells from the mouse EL4 (T lymphoma, ATCC TIB-39) cell line were stimulated (5 hr) with Phorbol 12-Myristate 13-Acetate (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 (containing Monensin) (Cat. No. 554724). The cells were washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BUV395 Rat IgG1 κ Isotype Control (Cat. No. 564059; Left Panel) or BD Horizon BUV395 Rat Anti-Mouse IL-17A antibody (Cat. No. 565246; Right Panel) using BD Biosciences' Protocol for Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis. Two-color contour plots showing the expression of IL-17A (or Ig Isotype control staining) versus Autofluorescence were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Flow cytometric analysis of IL-17A-producing mouse EL-4 cells. Cells from the mouse EL4 (T lymphoma, ATCC TIB-39) cell line were stimulated (5 hr) with Phorbol 12-Myristate 13-Acetate (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 (containing Monensin) (Cat. No. 554724). The cells were washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BUV395 Rat IgG1 κ Isotype Control (Cat. No. 564059; Left Panel) or BD Horizon BUV395 Rat Anti-Mouse IL-17A antibody (Cat. No. 565246; Right Panel) using BD Biosciences' Protocol for Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis. Two-color contour plots showing the expression of IL-17A (or Ig Isotype control staining) versus Autofluorescence were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
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BD Horizon™
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_2722575
Aqueous buffered solution containing BSA 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 with BD Horizon BUV395 under optimum conditions, and unconjugated antibody and free BD Horizon BUV395 were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 8,575,303; 8,354,239.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565246 Rev. 2
Antibody Details
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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.

The antibody was conjugated to BD Horizon BUV395 which has been exclusively developed by BD Biosciences as an optimal dye for use on a 355 nm laser equipped instrument. With an Ex Max at 348 nm  and an Em Max at 395 nm, this dye has virtually no spillover into any other detector. BD Horizon BUV395 can be excited with a 355 nm laser and detected with a 379/28 filter.

565246 Rev. 2
Format Details
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BUV395
The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV395
Ultraviolet 355 nm
348 nm
395 nm
565246 Rev.2
Citations & References
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Development References (4)

  1. Coquet JM, Chakravarti S, Kyparissoudis K, et al. Diverse cytokine production by NKT cell subsets and identification of an IL-17-producing CD4-NK1.1- NKT cell population. Proc Natl Acad Sci U S A. 2008; 105(32):11287-11292. (Clone-specific: Flow cytometry). View Reference
  2. Dong C. Th17 cells: Current understanding of their generation and regulation. Eur J Immunol. 2009; 39(3):640-644. (Biology). View Reference
  3. Lampropoulou V, Hoehlig K, Roch T, et al. TLR-activated B cells suppress T cell-mediated autoimmunity. J Immunol. 2008; 180(7):4763-4773. (Clone-specific: ELISA). View Reference
  4. 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
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565246 Rev. 2

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