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BD Horizon™ BV605 Rat Anti-Mouse IL-17A
Clone TC11-18H10 (RUO)
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), preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722).
The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BV605 Rat IgG1 κ Isotype Control (Cat. No. 562993; Left Panel) or BD Horizon BV605 Rat Anti-Mouse IL-17A (Cat. No. 564169; Right Panel) using BD Biosciences' Protocol for Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis. Two-color contour plots showing correlated expression patterns of IL-17A antibody (or Ig Isotype control staining) versus cellular 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™ LSR II Flow Cytometer 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), preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722).
The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BV605 Rat IgG1 κ Isotype Control (Cat. No. 562993; Left Panel) or BD Horizon BV605 Rat Anti-Mouse IL-17A (Cat. No. 564169; Right Panel) using BD Biosciences' Protocol for Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis. Two-color contour plots showing correlated expression patterns of IL-17A antibody (or Ig Isotype control staining) versus cellular 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™ LSR II Flow Cytometer 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), preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722).
The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BV605 Rat IgG1 κ Isotype Control (Cat. No. 562993; Left Panel) or BD Horizon BV605 Rat Anti-Mouse IL-17A (Cat. No. 564169; Right Panel) using BD Biosciences' Protocol for Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis. Two-color contour plots showing correlated expression patterns of IL-17A antibody (or Ig Isotype control staining) versus cellular 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™ LSR II Flow Cytometer 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), preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722).
The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BV605 Rat IgG1 κ Isotype Control (Cat. No. 562993; Left Panel) or BD Horizon BV605 Rat Anti-Mouse IL-17A (Cat. No. 564169; Right Panel) using BD Biosciences' Protocol for Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis. Two-color contour plots showing correlated expression patterns of IL-17A antibody (or Ig Isotype control staining) versus cellular 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™ LSR II Flow Cytometer System.
Product Details
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_2738640
Aqueous buffered solution containing ≤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™ BV605 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV605 were removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
- CF™ is a trademark of Biotium, Inc.
- 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
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.1 mg
Cat No.
553141
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Fixation and Permeabilization Solution RUO
Size
125 ML
Cat No.
554722
Brilliant Stain Buffer RUO
Size
100 Tests
Cat No.
563794
Perm/Wash Buffer RUO
Size
100 ML
Cat No.
554723
564169 Rev. 3
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.
This antibody is conjugated to BD Horizon BV605 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max of 602-nm, BD Horizon BV605 can be excited by a violet laser and detected with a standard 610/20-nm filter set. BD Horizon BV605 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em max at 605-nm. Due to the excitation of the acceptor dye by the green (532 nm) and yellow-green (561 nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers. BD Horizon BV605 conjugates are very bright, often exhibiting brightness equivalent to PE conjugates and can be used as a third color off of the violet laser.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).
564169 Rev. 3
Format Details
BV605
The BD Horizon Brilliant Violet™ 605 (BV605) dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 407-nm and an acceptor dye with an emission maximum (Em Max) at 605-nm. BV605, driven by BD innovation, is designed to be excited by the Violet laser (405-nm) and detected using an optical filter centered near 610-nm (e.g., a 610/20-nm bandpass filter). The acceptor dye can be excited by the Yellow-Green (561-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BV605
407 nm
605 nm
564169 Rev.3
Citations & References
View product citations for antibody "564169" on CiteAb
Development References (4)
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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
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Dong C. Th17 cells: Current understanding of their generation and regulation. Eur J Immunol. 2009; 39(3):640-644. (Biology). View Reference
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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: Flow cytometry). View Reference
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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
564169 Rev. 3
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.