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Purified Rat Anti-Mouse CD184
Purified Rat Anti-Mouse CD184
Flow cytometric analysis of CD184 expression on mouse thymocytes. BALB/c thymocytes were costained with Purified Rat anti-Mouse CD184 (Cat. No. 551852) at 0.5 µg/test and APC Rat anti-Mouse CD4 (Cat. No. 553051), followed by Biotin Mouse Anti-Rat IgG2b (Cat. No. 553898) and PE Streptavidin (Cat. No. 554061). The quadrant markers for the bivariate dot plot were set based on staining with Purified Rat IgG2b, κ Isotype Control (Cat. No. 553986; data not shown). Flow cytometry was performed on a FACScan™ system.
Flow cytometric analysis of CD184 expression on mouse thymocytes. BALB/c thymocytes were costained with Purified Rat anti-Mouse CD184 (Cat. No. 551852) at 0.5 µg/test and APC Rat anti-Mouse CD4 (Cat. No. 553051), followed by Biotin Mouse Anti-Rat IgG2b (Cat. No. 553898) and PE Streptavidin (Cat. No. 554061). The quadrant markers for the bivariate dot plot were set based on staining with Purified Rat IgG2b, κ Isotype Control (Cat. No. 553986; data not shown). Flow cytometry was performed on a FACScan™ system.
Product Details
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BD Pharmingen™
CXCR4, C-X-C chemokine receptor type 4; Fusin; LESTR; PB-CKR; Sdf1r
Mouse (QC Testing)
Rat IgG2b, κ
GST-NCXCR4 fusion protein
Flow cytometry (Routinely Tested), Western blot (Reported)
0.5 mg/ml
12767
AB_394273
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

The Purified Rat Anti-Mouse CD184 antibody can be used for the immunofluorscent staining and flow cytometric analysis of mouse leukocytes and cell lines that express CXCR4 (see figure). A multistep staining procedure is recommended to amplify immunofluorescent signals for the flow cytometric analysis of mouse CXCR4 expression:

Step 1: Incubate 10e6 cells with 0.5 µg of Purified Rat Anti-Mouse CD184 antibody at 4°C for 15-20 minutes. Wash cells two times with staining medium containing sodium azide (e.g., Dulbecco's' PBS or tissue culture medium [without phenol red and biotin] with 0.09% sodium azide and 2% heat-inactivated FCS or 0.2% BSA).

Step 2: Incubate the cells with Biotin Mouse Anti-Rat IgG2b (Cat. No. 553898) at 4°C for 20 minutes. Wash cells two times.

Step 3: Incubate the cells with ≤0.06 µg of PE Streptavidin (Cat. No. 554061) at 4°C for 20 minutes. Wash two times. Resuspend cells in staining medium and analyze stained cells by flow cytometry.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
551852 Rev. 3
Antibody Details
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2B11/CXCR4

The 2B11/CXCR4 monoclonal antibody specifically binds to mouse CD184, which is also known as the C-X-C Chemokine Receptor type 4 , CXCR4.  CXCR4 (previously known as Fusin and LESTR), is a seven-transmembrane, G-protein-coupled receptor. It is the specific receptor for the CXC chemokine, SDF-1/CXCL12.  Mouse CXCR4 shows 91% homology at the amino acid level with human CXCR4. CXCR4 is widely expressed by hematopoietic and non-hematopoietic cell types including neutrophils, monocytes, T cells, B cells, CD34-positive progenitor cells, endothelial cells, neurons and astrocytes.  Human CXCR4 is used by T-tropic HIV-1 as a co-receptor for viral entry.  The mouse Cxcr4 gene has been mapped to chromosome 1.

551852 Rev. 3
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
551852 Rev.3
Citations & References
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Development References (11)

  1. Bleul CC, Farzan M, Choe H, et al. The lymphocyte chemoattractant SDF-1 is a ligand for LESTR/fusin and blocks HIV-1 entry. Nature. 1996; 382(6594):829-833. (Biology). View Reference
  2. Bleul CC, Wu L, Hoxie JA, Springer TA, Mackay CR. The HIV coreceptors CXCR4 and CCR5 are differentially expressed and regulated on human T lymphocytes.. Proc Natl Acad Sci U S A. 1997; 94(5):1925-1930. (Biology). View Reference
  3. De La Luz Sierra M, Gasperini P, McCormick P, Zhu J, Tosato G. Transcription factor Gfi-1 induced by G-CSF is a negative regulator of CXCR4 in myeloid cells. Blood. 2007; 110(7):2276-2285. (Methodology: Western blot). View Reference
  4. Feng Y, Broder CC, Kennedy PE, Berger EA. HIV-1 entry cofactor: functional cDNA cloning of a seven-transmembrane, G protein-coupled receptor. Science. 1996; 272(5263):872-877. (Biology). View Reference
  5. Forster R, Kremmer E, Schubel A, et al. Intracellular and surface expression of the HIV-1 coreceptor CXCR4/fusin on various leukocyte subsets: rapid internalization and recycling upon activation. J Immunol. 1998; 160(3):1522-1531. (Immunogen). View Reference
  6. Gupta SK, Lysko PG, Pillarisetti K, Ohlstein E, Stadel JM. Chemokine receptors in human endothelial cells. Functional expression of CXCR4 and its transcriptional regulation by inflammatory cytokines. J Biol Chem. 1998; 273(7):4282-4287. (Biology). View Reference
  7. Heesen M, Berman MA, Benson JD, Gerard C, Dorf ME. Cloning of the mouse fusin gene, homologue to a human HIV-1 co-factor. J Immunol. 1996; 157(12):5455-5460. (Biology). View Reference
  8. Hesselgesser J, Halks-Miller M, DelVecchio V, et al. CD4-independent association between HIV-1 gp120 and CXCR4: functional chemokine receptors are expressed in human neurons. Curr Biol. 1997; 7(2):112-121. (Biology). View Reference
  9. Loetscher M, Geiser T, O'Reilly T, Zwahlen R, Baggiolini M, Moser B. Cloning of a human seven-transmembrane domain receptor, LESTR, that is highly expressed in leukocytes. J Biol Chem. 1994; 269(1):232-237. (Biology). View Reference
  10. Oberlin E, Amara A, Bachelerie F, et al. The CXC chemokine SDF-1 is the ligand for LESTR/fusin and prevents infection by T-cell-line-adapted HIV-1. Nature. 1996; 382(6594):833-835. (Biology). View Reference
  11. Schabath R, Muller G, Schubel A, Kremmer E, Lipp M, Forster R. The murine chemokine receptor CXCR4 is tightly regulated during T cell development and activation. J Leukoc Biol. 1999; 66(6):996-1004. (Biology). View Reference
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551852 Rev. 3

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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.