Description
The 2B8 monoclonal antibody specifically binds to CD117 (c-Kit), a transmembrane tyrosine-kinase receptor that is encoded by the Kit gene (formerly dominant white spotting, W). The c-Kit ligand (also known as steel factor, stem cell factor, and mast cell growth factor) encoded by the Kit1 gene (formerly steel, SI), is a co-mitogen for hematopoietic stem cells, myeloerythroid progenitors and a mast-cell differentiation factor. The KitW and Kit1SI mutant alleles have similar pleiotropic effects on the development of melanocytes, germ cells, and the hematopoietic system. In the adult bone marrow, CD117 is expressed on hematopoietic progenitor cells, including CD90 (Thy-1) low, TER-119-, CD45R/B220-, CD11b (Mac-1)-, Ly-6G (Gr-1)-, CD4-, CD8-, and Sca-1 (Ly-6A/E)+ multipotent hemotopoietic stem cells, progenitors committed to myeliod and/or erythroid lineages, and precursors of B and T lymphocytes. This widespread expression of CD117 in hematopoietic precursors is consistent with the participation of c-Kit and its ligand in the regulation of several hematopoietic lineages. Intrathymic expression of c-Kit and c-Kit ligand suggest that CD117 is also involved in the regulation of some events during the development of T lymphocytes. CD117 is also expressed by mast cells and by dendritic cells found in the periarteriolar lymphocytoc sheaths (T-cell areas) of splenic white pulp. The mAb 2B8 reportedly does not block the action of c-Kit. This clone 2B8 had been reported to cross-react with rat.
The antibody was conjugated to BD Horizon BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes. It is a polymer-based tandem dye developed exclusively by BD Biosciences. With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.
Format
- Format
BB700
- Excitation Source
Blue 488 nm
- Excitation Max
485 nm
- Emission Max
693 nm
BD Horizon Brilliant™ Blue 700 (BB700) is a dye that was exclusively developed by BD Biosciences as brighter alternative to PerCP-Cy5.5. This dye also has less cross laser excitation off the 405 nm laser, resulting in less spillover into the violet channels compared to PerCP-Cy5.5. Due to similar excitation and emission properties, BD Horizon BB700 and PerCP-Cy5.5 cannot be used simultaneously.
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 BD Horizon™ BB700 under optimum conditions, and unconjugated antibody and free BD Horizon BB700 were removed.
Product Notices
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- 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.
- The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
- 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.
- 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.
- BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
- Cy is a trademark of GE Healthcare.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes 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 or 566349).
When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.
For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
