BUV737 Mouse Anti-Human MIC A/B
Clone 6D4 (RUO)
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- Alternative Name MHC class I polypeptide-related sequence A/B; PERB11.1/ PERB11.2
- Concentration 0.2 mg/ml
- Isotype Mouse IgG2a, κ
- Reactivity Human (Tested in Development)
Flow cytometry (Qualified)
- Immunogen MICA transfected cells
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The 6D4 monoclonal antibody specifically binds to the human MHC class I polypeptide-related sequence A (MICA, aka PERB11.1) and B (MICB. aka PERB11.2) proteins. These ~70 kDa transmembrane glycoproteins are homologs of the major histocompatibility complex class I molecules although they lack association with β2 microglobulin. The MHC class I-related MICA and MICB chains are expressed by some gut epithelial cells in vivo. MICA and MICB expression by other epithelial cells and cell types, including fibroblasts and endothelial cells, is induced by stress, eg, stress caused by bacterial and viral infections, autoimmunity or cellular transformation. Epithelial cell expression of MICA and MICB has also been detected in transplanted kidneys and pancreas that show histological signs of rejection and or cellular injury. This suggests their potential role in transplant immunopathology. MICA and MICB are ligands for NKG2D (CD314), an activating receptor expressed by natural killer (NK) cells, γδ T cells, CD8+ and some CD4+ αβ T cells. The 6D4 antibody reportedly blocks NKG2D-positive NK cell- and T cell-mediated cytotoxicity against MICA/B-positive target cells.
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).