RB780 Mouse Anti-Human IL-17A

BD Horizon™ RB780 Mouse Anti-Human IL-17A

Name: RB780 Mouse Anti-Human IL-17A
Brand: BD Horizon™
SKU: 568765

Clone N49-653

(RUO)

Two-color flow cytometric analysis of IL-17A expression in stimulated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate (Sigma P-8139; 50 ng/ml final concentration) and Ionomycin (Sigma I-0634; 1 μg/ml final concentration) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) [Cat. No. 554724]. The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656], fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then washed with and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with APC Mouse Anti-Human CD4 antibody (Cat. No. 555349/561840/561841) and with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; Left Plot) or BD Horizon™ RB780 Mouse Anti-Human IL-17A antibody (Cat. No. 568765/568766; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of IL-17A (or Ig Isotype control staining) versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software.

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Product Details

Brand: BD Horizon™

Alternative Name: IL17; IL-17; CTLA8; Cytotoxic T-lymphocyte-associated serine esterase 8

Reactivity: Human (QC Testing)

Isotype: Mouse IgG1, κ

Immunogen: Human IL-17A Recombinant Protein

Application: Intracellular staining (flow cytometry) (Routinely Tested)

Vol. Per Test: 5 µl

Entrez Gene ID: 3605

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: 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 the dye under optimum conditions and unreacted dye was removed.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
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.
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.
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 http://regdocs.bd.com to access safety data sheets (SDS).
Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Protein Transport Inhibitor (Containing Monensin) RUO

Size 0.7 mL Cat No. 554724

Fixation Buffer RUO

Size 100 mL Cat No. 554655

Perm/Wash Buffer RUO

Size 100 mL Cat No. 554723

RB780 Mouse IgG1, κ Isotype Control RUO

Size 50 µg Cat No. 568532

View Details

568765 Rev. 2

Antibody Details

N49-653

Human IL-17A, also known as IL-17, is a proinflammatory cytokine that is encoded by the IL17A gene in chromosome 6. IL-17A is produced as a disulfide-linked homodimer comprised of two mature 136-amino acid polypeptides. It is a member of the IL-17 family of structurally related cytokines, designated IL-17A through IL-17F. Activated memory T cells, especially Th17 cells (specialized IL-17A-producing CD4+ T cells distinct from Th1 and Th2 cells) produce IL-17 and provide protective immunity against pathogens. Activated CD8+ T cells, γδT cells, NK cells and neutrophils can also be activated to produce IL-17A. IL-17A binds to and exerts its biological activity through IL-17 receptors (IL-17R) that are expressed by a variety of target cells including fibroblasts, epithelial and endothelial cells, monocytes/macrophages and mast cells. The ubiquitous IL-17R expression pattern may explain the broad tissue responsiveness to IL-17. IL-17 induces stromal cells to secrete cytokines and chemokines involved in inflammatory and hematopoietic processes. For example, IL-17 induces fibroblasts to produce IL-6, IL-8, G-CSF and express increased surface ICAM-1. The N49-653 antibody reacts with human IL-17A.

568765 Rev. 2

Format Details

RB780

The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.

Format: RB780

Excitation Source: Blue 488 nm

Excitation Max: 498 nm

Emission Max: 781 nm

568765 Rev.2

Citations & References

Development References (9)

Fossiez F, Djossou O, Chomarat P, et al. T cell interleukin-17 induces stromal cells to produce proinflammatory and hematopoietic cytokines. J Exp Med. 1996; 183(6):2593-2603. (Biology). View Reference
Korn T, Oukka M, Kuchroo V, Bettelli E. Th17 cells: effector T cells with inflammatory properties. Semin Immunol. 2007; 19(6):362-371. (Biology). View Reference
Melton AC, Melrose J, Alajoki L, et al. Regulation of IL-17A production is distinct from IL-17F in a primary human cell co-culture model of T cell-mediated B cell activation. PLoS ONE. 2013; 8(3):e58966. (Clone-specific: Flow cytometry). View Reference
Moseley TA, Haudenschild DR, Rose L, Reddi AH. Interleukin-17 family and IL-17 receptors. Cytokine Growth Factor Rev. 2003; 14(2):155-174. (Biology). View Reference
Pennino D, Bhavsar PK, Effner R, et al. IL-22 suppresses IFN-gamma-mediated lung inflammation in asthmatic patients. J Allergy Clin Immunol. 2013; 131(2):562-570. (Clone-specific: Flow cytometry). View Reference
Weaver CT, Hatton RD, Mangan PR, Harrington LE. IL-17 family cytokines and the expanding diversity of effector T cell lineages. Annu Rev Immunol. 2007; 25:821-852. (Biology). View Reference
Yao Z, Painter SL, Fanslow WC, et al. Human IL-17: a novel cytokine derived from T cells. J Immunol. 1995; 155(12):5483-5486. (Biology). View Reference
Yao Z, Spriggs MK, Derry JM, et al. Molecular characterization of the human interleukin (IL)-17 receptor. Cytokine. 1997; 9(11):794-800. (Biology). View Reference
Zhao M, Tan Y, Peng Q, et al. IL-6/STAT3 pathway induced deficiency of RFX1 contributes to Th17-dependent autoimmune diseases via epigenetic regulation.. Nat Commun. 2018; 9(1):583. (Clone-specific: Flow cytometry). View Reference

View All (9)

568765 Rev. 2

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