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BD Horizon™ BB515 Mouse Anti-Human CCR8 (CD198)
Clone 433H (RUO)
Multiparameter flow cytometric analysis of CCR8 (CD198) on human peripheral blood leucocytes. Whole blood was stained with APC Mouse Anti-Human CD3 antibody (Cat. No. 555335/561810/561811) and either BD Horizon™ BB515 Mouse IgG2a, κ Isotype Control (Cat. No. 564515; Left Plots) or BD Horizon™ BB515 Mouse Anti-Human CCR8 (CD198) antibody (Cat. No. 566891/566892; Right Plots). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-parameter pseudocolor density plots showing the correlated expression of CCR8 (CD198) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals [Top Plots] or CD3 [Bottom Plots] were derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations or lymphocytes, respectively. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Multiparameter flow cytometric analysis of CCR8 (CD198) on human peripheral blood leucocytes. Whole blood was stained with APC Mouse Anti-Human CD3 antibody (Cat. No. 555335/561810/561811) and either BD Horizon™ BB515 Mouse IgG2a, κ Isotype Control (Cat. No. 564515; Left Plots) or BD Horizon™ BB515 Mouse Anti-Human CCR8 (CD198) antibody (Cat. No. 566891/566892; Right Plots). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-parameter pseudocolor density plots showing the correlated expression of CCR8 (CD198) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals [Top Plots] or CD3 [Bottom Plots] were derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations or lymphocytes, respectively. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Product Details
BD Horizon™
CCR8; CKR-L1; CKRL1; CMKBR8; CMKBRL2; CY6; GPR-CY6; GPRCY6; MGC129966; MGC1
Human (QC Testing)
Mouse BALB/c IgG2a, κ
Human CCR8 Transfected Cell Line
Flow cytometry (Routinely Tested)
5 µl
AB_2869939
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™ BB515 under optimum conditions and unconjugated antibody was removed.
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
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.
- 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.
- The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor® is a registered trademark of Life Technologies Corporation.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Brilliant Stain Buffer RUO
Size
100 Tests
Cat No.
563794
BB515 Mouse IgG2a, κ Isotype Control RUO
Size
50 µg
Cat No.
564515
BB515 Mouse Anti-Human CCR8 (CD198) RUO
Size
25 Tests
Cat No.
566892
APC Mouse Anti-Human CD3 RUO
Size
100 Tests
Cat No.
555335
566892 Rev. 1
Antibody Details
433H
The 433H monoclonal antibody specifically binds to C-C chemokine receptor type 8 (CC-CKR-8 or CCR8) which is also known as CD198, GPRCY6, CY6, CKRL1, CMKBR8, CMKBRL2, or TER1. CCR8 (CD198) is a seven-transmembrane, G-protein-coupled, glycoprotein receptor that belongs to the beta chemokine receptor family. CCR8 is expressed on monocytes, macrophages, eosinophils, Langerhans cells, thymocytes, and vascular smooth muscle cells. It is preferentially expressed on some T cell subsets including skin resident memory T cells, type 2 T-helper (Th2-like) cells, and T regulatory cells. CCR8 is a receptor for the chemokine C-C motif chemokine 1 (CCL1), also known as SCYA1 or I-309, and plays a role in regulating monocyte and T cell chemotaxis and thymocyte apoptosis. CCR8 may play a role in lung T cell recruitment in asthma and can serve as an alternative coreceptor to CD4 for HIV-1 infection. The 433H antibody reportedly neutralized the chemotactic response of cultured T cells to CCL1.
The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.
566892 Rev. 1
Format Details
BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye. It is not recommended to use BB515 in cocktails that include streptavidin conjugates as it may cause high background.
BB515
490 nm
515 nm
566892 Rev.1
Citations & References
View product citations for antibody "566892" on CiteAb
Development References (8)
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D'Ambrosio D, Iellem A, Bonecchi R, et al. Selective up-regulation of chemokine receptors CCR4 and CCR8 upon activation of polarized human type 2 Th cells.. J Immunol. 1998; 161(10):5111-5. (Biology). View Reference
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Fox JM, Najarro P, Smith GL, Struyf S, Proost P, Pease JE. Structure/function relationships of CCR8 agonists and antagonists. Amino-terminal extension of CCL1 by a single amino acid generates a partial agonist.. J Biol Chem. 2006; 281(48):36652-61. (Biology). View Reference
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Haque NS, Fallon JT, Pan JJ, Taubman MB, Harpel PC. Chemokine receptor-8 (CCR8) mediates human vascular smooth muscle cell chemotaxis and metalloproteinase-2 secretion.. Blood. 2004; 103(4):1296-304. (Biology). View Reference
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Lee S, Tiffany HL, King L, Murphy PM, Golding H, Zaitseva MB. CCR8 on human thymocytes functions as a human immunodeficiency virus type 1 coreceptor.. J Virol. 2000; 74(15):6946-52. (Biology). View Reference
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Louahed J, Struyf S, Demoulin JB, et al. CCR8-dependent activation of the RAS/MAPK pathway mediates anti-apoptotic activity of I-309/ CCL1 and vMIP-I.. Eur J Immunol. 2003; 33(2):494-501. (Biology). View Reference
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McCully ML, Ladell K, Hakobyan S, Mansel RE, Price DA, Moser B. Epidermis instructs skin homing receptor expression in human T cells.. Blood. 2012; 120(23):4591-8. (Clone-specific: Flow cytometry). View Reference
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Mutalithas K, Guillen C, Raport C, et al. Expression of CCR8 is increased in asthma.. Clin Exp Allergy. 2010; 40(8):1175-85. (Immunogen: Blocking, Flow cytometry, Functional assay, Immunohistochemistry, Inhibition). View Reference
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Pease JE. Is there a role for CCR8 in the pathogenesis of asthma?. Clin Exp Allergy. 2010; 40(8):1110-2. (Clone-specific). View Reference
566892 Rev. 1
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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.