PerCP-Cy™5.5 Mouse Anti-Human CD184
Clone 12G5 (RUO)
- Brand BD Pharmingen™
- Alternative Name CXCR4; Fusin; SDF-1 receptor; LAP3; LCR1; LESTR; NPYY3R; NPY3R; WHIM; HM89
- Vol. Per Test 5 µl
- Isotype Mouse BALB/c IgG2a, κ
- Reactivity Rhesus, Cynomolgus, Baboon (QC Testing)
Flow cytometry (Routinely Tested)
- Immunogen SIVmac variant CP-MAC-infected Sup-T1 cells
- Workshop No. VII 70204, 70305
- Entrez Gene ID 7852
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The 12G5 monoclonal antibody specifically binds to CD184, also known as CXCR4 and Fusin. CD184/CXCR4 is a seven-transmembrane domain, G-protein-linked, glycoprotein chemokine receptor. CD184 serves as a receptor for the C-X-C chemokine, SDF-1. It is expressed on a wide variety of hematopoietic cells including lymphoid and myeloid precursor cells, megakaryocytes, platelets, T and B lymphocytes, granulocytes, monocytes/macrophages, and dendritic cells. It is also expressed on vascular endothelial cells, epithelial cells, neurons and astrocytes. CD184 plays a variety of roles in hematopoiesis, vascularization and neural development. CD184 also functions as a coreceptor for infection with T-cell tropic strains of HIV-1 and as a receptor for CD4-independent infection by some HIV isolates. The 12G5 antibody has been reported to block CD4-independent infection by HIV-2 and CD4-dependent infection by some T-cell tropic isolates of HIV-1.
PerCP-Cy™5.5 is a tandem conjugate that combines PerCP with a cyanine dye. PerCP-Cy5.5 is not subject to photobeaching like PerCP and can be used with stream-in-air flow cytometers. It has less Fc receptor–mediated nonspecific staining than PE-Cy5. Additionally, the PerCP-Cy5.5 tandem conjugate is not as susceptible to fixative or light instability compared to APC-Cy7 and PE-Cy7.
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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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
- PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- 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.
- This PerCP-conjugated product is sold under license to the following patent: US Patent No. 4,876,190.
- Cy is a trademark of Amersham Biosciences Limited.
- This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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 alternative staining procedures may be neccessary. A multiple-step staining procedure is strongly recommended, in some instances, to amplify immunofluorescent signals for the flow cytometric analysis of CXCR4 expression. Investigators may find the Purified Mouse Anti-Human CD184 antibody (MN 555972) 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) or PerCP-Cy™5.5 Streptavidin (MN 551419).