Description
The C34-3448 monoclonal antibody specifically binds to CD195 which is also known as, C-C chemokine receptor type 5 (CCR5). CD195 is a seven transmembrane-spanning G-protein-coupled receptor that belongs to the β-chemokine receptor family. CD195 regulates lymphocyte chemotaxis activation and transendothelial migration during inflammation. It signals in response to at least three chemokines: CCL3/MIP-1α, CCL4/MIP-1β, and CCL5/RANTES. CD195 is expressed on macrophages and some T-lymphocytes.
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.
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.
1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
5. An isotype control should be used at the same concentration as the antibody of interest.
6. CF™ is a trademark of Biotium, Inc.
7. 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.
8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
9. For U.S. patents that may apply, see bd.com/patents.
10. 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.
11. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
12. Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
Specifications

General

SourceMouse B7-H1, amino acids (Phe19-Thr238) (Accession# NM_021893), was expressed in 293E cells. Human IgG-Fc is in the carboxy-terminus.
Molecular MassThe 458 amino acid recombinant protein has a predicted molecular mass of approximately 51.5kD. The DTT-reduced and non-reduced protein migrate at approximately 70 - 80kD and 140kD respectively by SDS-PAGE. The predicted N-terminal amino acid is Phe.
Purity>95%, as determined by Coomassie stained SDS-PAGE.
Formulation0.22 µm filtered protein solution is in PBS, pH 7.2.
Endotoxin LevelLess than 0.1 EU per µg of protein as determined by the LAL method.
Concentration10 and 25 µg sizes are bottled at 200 µg/mL. 100 µg size and larger sizes are lot-specific and bottled at the concentration indicated on the vial. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.
Storage & HandlingUnopened vial can be stored between 2°C and 8°C for up to 2 weeks, at -20°C for up to six months, or at -70°C or colder until the expiration date. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored at -20°C or colder. Stock solutions can also be prepared at 50 - 100 µg/mL in appropriate sterile buffer, carrier protein such as 0.2 - 1% BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to one week and stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.
ActivityImmobilized mouse PD-L1 binds mouse PD-1 in a dose dependent manner. The ED50 = 0.1 - 0.5 µg/ml.
Recommended UsageBioassay
Application NotesBioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are verified in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at [email protected].
Product CitationsWang H, et al. 2022. Mol Cancer Ther. 21:1710. PubMed Pereira JA, et al. 2023. Front Immunol. 14:997376. PubMed Smith BC, et al. 2023. J Neuroinflammation. 234:20. PubMed Cheng B, et al. 2022. Cancer Commun (Lond). 42:17. PubMed Mitchell LA, et al. 2019. Oncotarget. 10:2252. PubMed Liao YC, et al. 2021. Front Immunol. 12:743030. PubMed
StructureDimer.
DistributionAPC, monocytes, dendritic cells, expressed in nonlymphoid tissues, and stromal cell.
FunctionEnhances CD28-independent T-helper cell function, suppresses immune responses against autoantigens, and participates in fetomaternal tolerance.
InteractionAntigen-stimulated T and B cells, regulatory T cells, follicular T and B cells, dendritic cells, and monocytes.
Ligand/ReceptorPD-1 (CD279).
BioactivityImmobilized mouse PD-L1 binds to its receptor PD-1.
Cell TypeEmbryonic Stem Cells
Biology AreaImmunology, Cancer Biomarkers, Costimulatory Molecules, Stem Cells
Molecular FamilyAdhesion Molecules, CD Molecules, Immune Checkpoint Receptors, Soluble Receptors
Antigen References1. Dong H, et al. 1999. Nat. Med. 5:1365.2. Freeman GJ, et al. 2000. J. Exp. Med. 192:1027.3. Latchman Y, et al. 2001. Nat. Immunol. 2:261.4. Latchman YE, et al. 2004. Proc. Natl. Acad. Sci. USA 101:10691.5. Guleria I, et al. 2005. J. Exp. Med. 202:231.6. Lin DY, et al. 2008. Proc. Natl. Acad. Sci. USA 105:3011.7. Dai S, et al. 2014. Cell Immunol. 290:72.8. Melero I, et al. 2015. Nat. Rev. Cancer 15:457.
Gene ID60533
UniProtView information about B7-H1 on UniProt.org
Regulatory StatusRUO
Other NamesCD274, Programmed cell death 1 ligand 1 (PD-L1), B7 homolog 1 (B7h1)
BD 758206 RB613 Rat Anti-Mouse CD195 (CCR5) C34-3448.1.1 | IRIGHT