Description
The 2G8 monoclonal antibody specifically binds to the mouse C-X-C Chemokine Receptor type 5, CXCR5. CXCR5 is also known as CD185, BLR1, NLR and MDR15. CXCR5 is a seven-transmembrane, G-protein-coupled receptor that is specific for the CXC chemokine, CXCL13/BLC/BCA-1. The expression of CXCR5 has been detected in spleen, lymph nodes, tonsils, brain, bone marrow, T cells, B cells, cerebrum, cerebellum, hippcampus and pituitary. In mouse spleen, CXCR5 was strictly expressed by mature B cells and a small subset of T lymphocytes. CXCR5 plays a role in directing the migration of B and T cells to B cell follicles with the spleen and certain other lymphoid tissues. The immunogen used to generate 2G8 hybridoma was a recombinant protein containing N-terminal amino acids of mouse CXCR5 (GST-NmBLR1).
Format
- Format
PerCP-Cy™5.5
- Excitation Source
Blue 488 nm
- Excitation Max
482 nm
- Emission Max
678 nm
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.
Suggested Companion Products
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.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- 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.
- 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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Cy is a trademark of GE Healthcare.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- 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 mouse CXCR5 expression. Investigators may find the Purified Rat Anti-Mouse CXCR5 antibody (MN 551961) to be useful in conjunction with appropriate secondary and tertiary reagents for detecting low frequency expression, such as with Biotin Mouse Anti-Rat IgG2a (MN 553894) and PE Streptavidin (MN 554061) or
PerCP-Cy™5.5 Streptavidin (MN 551419).
