BUV737 Mouse Anti-Human CD49d
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- Concentration 0.2 mg/ml
- Isotype Mouse BALB/c IgG2b, κ
- Reactivity Human (Tested in Development)
Flow cytometry (Qualified)
- Immunogen CD8+ T-cell Line
- Workshop No. V BP BP378, AS S216
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
Regulatory Status Legend
The L25 monoclonal antibody specifically recognizes CD49d which is also known as Integrin alpha-4 (α4 Integrin) or the alpha chain of the Very-Late Antigen-4 (VLA-4α). CD49d is a ~150 kDa type I transmembrane glycoprotein that is encoded by ITGA4 and belongs to the integrin family of cell adhesion molecules. VLA-4, like other integrins, is a noncovalently associated heterodimeric glycoprotein composed of á and â subunits and is involved in cell-cell and cell-extracellular matrix interactions. The â chain of the VLA-4 complex is the CD29 antigen, a 130 kDa glycoprotein. The CD29 antigen, also known as the â1 subunit, is common to the VLA family of integrins. When acting as a matrix receptor, the CD49d antigen binds to CS-1, an alternatively spliced domain of fibronectin. When functioning as a cell receptor, the CD49d antigen binds to the vascular cell-adhesion molecule-1 (VCAM-1). The interaction between the CD49d antigen and VCAM-1 is known to play an important role in stabilizing the adhesion of lymphocytes to endothelial cells and in mediating B-lymphocyte precursor/bone marrow stromal cell adhesion. The CD49d antigen, when associated with the β7 integrin, forms the α4/β7 integrin lymphocyte homing receptor for Peyer's patches, binding to the mucosal vascular addressin MAdCAM-1. The CD49d antigen is also involved in CD3-dependent CD4+ T-lymphocyte activation via its interaction with fibronectin. The CD49d antigen is primarily expressed on T and B lymphocytes and weakly expressed on monocytes. The L25 antibody can block or enhance fibronectin-stimulated T-lymphocyte proliferation. It immunoprecipitates three proteins of 150 kDa, 85 kDa, and 75 kDa under both reducing and nonreducing conditions from HPB-ALL cells, B lymphoblasts, peripheral blood lymphocytes, and IL-2-dependent cell lines.
The antibody was conjugated to BD Horizon™ BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 737-nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 filter. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into channels detecting Alexa Fluor® 700-like dyes (eg, 712/20-nm filter).
Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV737 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV737 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.
BUV737 is a tandem fluorochrome that combines BD Horizon BUV395 and an acceptor dye with an Em Max at 737 nm. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover in to channels detecting Alexa Fluor® 700 like dyes (for example, 712/20-nm filter). BUV737 has been exclusively developed by BD Biosciences for instruments equipped with a 355-nm UV laser.
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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 BD Horizon BUV737 under optimal conditions that minimize unconjugated dye and antibody.
- This antibody was developed for use in flow cytometry.
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- 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.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Ultraviolet 737 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).