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Purified Mouse Anti-Human CD194
Purified Mouse Anti-Human CD194

Flow cytometric detection of CD194 (CCR4) on human peripheral lymphocytes.  Human PBMC were stained with 0.25 µg of the purified Mouse Anti-Human CD194 (CCR4) antibody using a Biotin Goat Anti-Mouse Ig secondary antibody and with PE Streptavidin in conjunction with either a FITC Mouse Anti-Human CD4 antibody (Cat. No. 555346, left panel) or with a FITC Mouse Anti-Human CD8 antibody (Cat. No. 555366, right panel). The data reflects gating on lymphocytes, based on forward and side scattered light signals. The level of nonspecific staining was assessed by using purified mouse IgG1 (Cat. No. 555746) as an isotype control. The quadrant markers for the bivariate dot plots were set based on the isotype control.

Flow cytometric detection of CD194 (CCR4) on human peripheral lymphocytes.  Human PBMC were stained with 0.25 µg of the purified Mouse Anti-Human CD194 (CCR4) antibody using a Biotin Goat Anti-Mouse Ig secondary antibody and with PE Streptavidin in conjunction with either a FITC Mouse Anti-Human CD4 antibody (Cat. No. 555346, left panel) or with a FITC Mouse Anti-Human CD8 antibody (Cat. No. 555366, right panel). The data reflects gating on lymphocytes, based on forward and side scattered light signals. The level of nonspecific staining was assessed by using purified mouse IgG1 (Cat. No. 555746) as an isotype control. The quadrant markers for the bivariate dot plots were set based on the isotype control.

Product Details
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BD Pharmingen™
CCR4; C-C chemokine receptor type 4; CMKBR4; K5-5
Human (QC Testing)
Mouse C57BL/6 IgG1, κ
Human CCR4 Transfected Cell Line
Flow cytometry (Routinely Tested)
0.5 mg/ml
HCDM 2006
1233
AB_2074502
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Flow cytometry:  Chemokine receptors are known to internalize during manipulation resulting in low frequency expression.  Investigators are advised to perform immunophenotyping studies of chemokine receptors on freshly collected samples (<24 Hrs). Incubation with the antibody should be done at 4°C in the dark. Cellular manipulation, such as Ficoll separation, freezing, or exposure to cold temperatures prior to staining should be minimized and have been shown to cause a decrease in staining intensity and/or inconsistent results.

Investigators should note that a multiple-step staining procedure is recommended, in some instances, to amplify immunofluorescent signals for the flow cytometric analysis of CCR4 expression.  Investigators may find the Purified Mouse Anti-Human CD194 (CCR4) antibody (MN 551121) to be useful in conjunction with appropriate secondary and tertiary reagents for detecting low frequency expression, such as with Biotin Goat Anti-Mouse Ig (MN 553999) and PE Streptavidin (MN 554061).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. 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.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
551121 Rev. 2
Antibody Details
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1G1

The 1G1 monoclonal antibody specifically binds to CD194, also known as the human CC Chemokine Receptor type 4 (CCR4).  CCR4 is expressed on activated Th2 cells, regulatory T cells, activated NK cells, basophils, monocytes and platelets.  CCR4 is a seven-transmembrane, G-protein-coupled receptor, and is the specific receptor for CC chemokines, CCL22/MDC/Macrophage-Derived Chemokine and CCL17/TARC/Thymus and Activation-Regulated Chemokine. It has been reported that CCR4 mRNA is expressed mainly in the thymus and spleen. The human CCR4 gene has been mapped to chromosome 3p24. The purified form of this antibody has been reported not to be a neutralizing antibody.  The immunogen used to generate the 1G1 hybridoma has been reported to be human CCR4 transfected L1.2 mouse lymphoma cells.

551121 Rev. 2
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
551121 Rev.2
Citations & References
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Development References (9)

  1. Andrew DP, Ruffing N, Kim CH, et al. C-C chemokine receptor 4 expression defines a major subset of circulating nonintestinal memory T cells of both Th1 and Th2 potential. J Immunol. 2000; 166(1):103-111. (Biology). View Reference
  2. Bonecchi R, Bianchi G, Bordignon PP, et al. Differential expression of chemokine receptors and chemotactic responsiveness of type 1 T helper cells (Th1s) and Th2s. J Exp Med. 1998; 187(1):129-134. (Biology). View Reference
  3. Campbell JJ, Haraldsen G, Pan J, et al. The chemokine receptor CCR4 in vascular recognition by cutaneous but not intestinal memory T cells. Nature. 1999; 400(6746):776-780. (Biology). View Reference
  4. 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
  5. Imai T, Baba M, Nishimura M, Kakizaki M, Takagi S, Yoshie O. The T cell-directed CC chemokine TARC is a highly specific biological ligand for CC chemokine receptor 4. J Biol Chem. 1997; 272(23):15036-15042. (Biology). View Reference
  6. Imai T, Chantry D, Raport CJ, et al. Macrophage-derived chemokine is a functional ligand for the CC chemokine receptor 4. J Biol Chem. 1998; 273(3):1764-1768. (Biology). View Reference
  7. Power CA, Meyer A, Nemeth K, et al. Molecular cloning and functional expression of a novel CC chemokine receptor cDNA from a human basophilic cell line. J Biol Chem. 1995; 270(33):19495-19500. (Biology). View Reference
  8. Sallusto F, Lenig D, Mackay CR, Lanzavecchia A. Flexible programs of chemokine receptor expression on human polarized T helper 1 and 2 lymphocytes. J Exp Med. 1998; 187(6):875-883. (Biology). View Reference
  9. Samson M, Soularue P, Vassart G, Parmentier M. The genes encoding the human CC-chemokine receptors CC-CKR1 to CC-CKR5 (CMKBR1-CMKBR5) are clustered in the p21.3-p24 region of chromosome 3. Genomics. 1996; 36(3):522-526. (Biology). View Reference
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551121 Rev. 2

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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.