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PE Mouse Anti-Human CCR8 (CD198)
PE Mouse Anti-Human CCR8 (CD198)
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 PE Mouse IgG2a, κ Isotype Control (Cat. No. 553457; Left Plots) or PE Mouse Anti-Human CCR8 (CD198) antibody (Cat. No. 566899/566900; 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 PE Mouse IgG2a, κ Isotype Control (Cat. No. 553457; Left Plots) or PE Mouse Anti-Human CCR8 (CD198) antibody (Cat. No. 566899/566900; 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
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BD Pharmingen™
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_2869942
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 R-PE under optimum conditions, and unconjugated antibody and free PE were 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 CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566899 Rev. 4
Antibody Details
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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.

        

566899 Rev. 4
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
566899 Rev.4
Citations & References
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Development References (8)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
View All (8) View Less
566899 Rev. 4

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.