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RB780 Mouse Anti-Human CD194
RB780 Mouse Anti-Human CD194
Multiparameter flow cytometric analysis using BD OptiBuild™ RB780 Mouse Anti-Human CD194 antibody (Cat. No. 755494) on human peripheral blood. Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Multiparameter flow cytometric analysis using BD OptiBuild™ RB780 Mouse Anti-Human CD194 antibody (Cat. No. 755494) on human peripheral blood. Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Product Details
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BD OptiBuild™
CCR4; C-C chemokine receptor type 4; CMKBR4; K5-5
Human (Tested in Development)
Mouse C57BL/6 IgG1, κ
Human CCR4 Transfected Cell Line
Flow cytometry (Qualified)
0.2 mg/ml
HCDM 2006
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 to the dye under optimum conditions that minimize unconjugated dye and antibody.

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

  1. 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.
  2. Researchers should determine the optimal concentration of this reagent for their individual applications.
  3. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. 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.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. 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.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  12. 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.
755494 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.

755494 Rev. 2
Format Details
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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.
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RB780
Blue 488 nm
498 nm
781 nm
755494 Rev.2
Citations & References
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View product citations for antibody "755494" on CiteAb

Development References (11)

  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. (Immunogen: Blocking, Flow cytometry, Functional assay, Inhibition). 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. Imai T, Nagira M, Takagi S, et al. Selective recruitment of CCR4-bearing Th2 cells toward antigen-presenting cells by the CC chemokines thymus and activation-regulated chemokine and macrophage-derived chemokine. Int Immunol. 1999; 11(1):81-88. (Biology). View Reference
  8. 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
  9. 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
  10. 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
  11. Zola H, Swart B, Banham A, et al. CD molecules 2006--human cell differentiation molecules.. J Immunol Methods. 2007; 319(1-2):1-5. (Clone-specific). View Reference
View All (11) View Less
755494 Rev. 2

 

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