The L230 monoclonal antibody specifically recognizes CD51, which is also known as Integrin αV. CD51 (Integrin αV) is an ~150 kDa type I integral membrane glycoprotein that is encoded by ITGAV (integrin subunit alpha V) which belongs to the αV integrin subfamily. Integrin αV is cleaved into disulfide-linked 125 kDa and 25 kDa subunits. CD51 (Integrin αV) forms noncovalent heterodimeric complexes with five different integrin β chains: β1 (CD29), β3 (CD61), β5, β6, or β8. These heterodimeric integrins are variably expressed on different cell types including platelets, fibroblasts, endothelial cells, epithelial cells, monocytes, macrophages, dendritic cells, neutrophils, NK cells, osteoclasts, T cells, and B cells as well as pancreatic beta cells and tumor cells. These cell surface receptors bind to extracellular matrix and cell surface proteins including fibronectin, fibrinogen, vitronectin, thrombospondin, von Willebrand factor, laminin, bone sialoprotein, and osteopontin that contain an RGD (arginine-glycine-aspartic acid) motif. Other ligands include CD31 (PECAM-1), that binds to CD51/CD61 (αVβ3, vitronectin receptor) and is involved in transendothelial leukocyte migration to inflammatory sites, as well as cytokines and growth factors like latent TGF-β. These receptors play central roles in cellular adhesion and migration, as well as in the growth, proliferation, differentiation and survival of normal and tumor cells. The L230 antibody can reportedly block CD51 (Integrin αV) functions.
Bead-based compensation or unmixing controls, such as BD® CompBeads or BD™ SpectraComp™, can be used as surrogates to assess fluorescence spillover when bound to fluorochrome-conjugated antibodies. Although these beads have spectral properties similar to cells, variations in spectral emission may occur, resulting in differing spillover values compared to biological controls. Therefore, it is considered best practice to compare the spillover obtained from cells and bead-based controls when using BD® CompBeads or BD™ SpectraComp™ for the first time, to ensure they are appropriate for the intended 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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
3. For U.S. patents that may apply, see bd.com/patents.
4. 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.
5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
6. 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.
7. 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.
8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
9. An isotype control should be used at the same concentration as the antibody of interest.
10. 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.
11. Alexa Fluor™ is a trademark of Life Technologies Corporation.
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.
