Two-color flow cytometric analysis of CD196 (CCR6) expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PE Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553089/553090/561878) and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left Panel) or BD Horizon BV421Rat Anti-Mouse CD196 (CCR6) antibody (Cat. No. 564736; Right Panel). Two-color flow cytometric contour plots showing the correlated expression of CD196 (CCR6) [or Ig Isotype control staining] versus CD45R/B220 were derived from gated events with the forward and side light- scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of CD196 (CCR6) expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PE Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553089/553090/561878) and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left Panel) or BD Horizon BV421Rat Anti-Mouse CD196 (CCR6) antibody (Cat. No. 564736; Right Panel). Two-color flow cytometric contour plots showing the correlated expression of CD196 (CCR6) [or Ig Isotype control staining] versus CD45R/B220 were derived from gated events with the forward and side light- scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
BD Horizon™
CCR6, BN-1, CKR-L3, DRY6, KY411, GPR-CY4, STRL22
Mouse (QC Testing)
Rat IgG2a
Mouse CCR6-transfected Rat Cell Line
Flow cytometry (Routinely Tested)
5 µl
12458
AB_2738926
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 BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- Brilliant Violet™ 421 is a trademark of Sirigen.
- 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
0.1 mg
Cat No.
553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Brilliant Stain Buffer RUO
Size
100 Tests
Cat No.
563794
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
564736 Rev. 1
Antibody Details
140706
The 140706 monoclonal antibody specifically binds to mouse CD196 (CCR6), a CC chemokine receptor that is expressed on B lymphocytes and subsets of T and dendritic cells. The known ligand for CCR6 is CCL20/MIP-3α, and together they regulate the migration and recruitment of antigen-presenting and immunocompetent cells during inflammatory and immunological responses.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
564736 Rev. 1
Format Details
BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BV421
Violet 405 nm
407 nm
423 nm
564736 Rev.1
Citations & References
Development References (3)
-
Hirota K, Yoshitomi H, Hashimoto M, et al. Preferential recruitment of CCR6-expressing Th17 cells to inflamed joints via CCL20 in rheumatoid arthritis and its animal model. J Exp Med. 2007; 204(12):2803-2812. (Clone-specific: Flow cytometry). View Reference
-
Kucharzik T, Hudson JT 3rd, Waikel RL, Martin WD, Williams IR. CCR6 expression distinguishes mouse myeloid and lymphoid dendritic cell subsets: demonstration using a CCR6 EGFP knock-in mouse. Eur J Immunol. 2002; 32(1):104-112. (Biology). View Reference
-
Varona R, Villares R, Carramolino L, et al. CCR6-deficient mice have impaired leukocyte homeostasis and altered contact hypersensitivity and delayed-type hypersensitivity responses. J Clin Invest. 2001; 107(6):R37-R45. (Biology). View Reference
564736 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.
