Description: The 2B8 monoclonal antibody reacts with mouse CD117, also known as c-Kit receptor, Steel factor receptor, and stem cell factor receptor. A member of the tyrosine kinase receptor family, this 145-kDa molecule is expressed by a majority of hematopoietic progenitor cells characterized in the mouse bone marrow as a small subset of cells positive for Sca-1 and Thy1 (Thy1 low) and negative for lineage markers. The interaction of the mouse c-Kit receptor and steel factor promotes the proliferation and differentiation of hematopoietic progenitor cells. CD117 is also expressed by mast cells and plays a role in signaling and activation of these cells.
Applications Reported: This 2B8 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This 2B8 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. This can be used at less than or equal to 0.125 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Super Bright 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.
When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.
In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497).
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 405 nm; Emission: 780 nm; Laser: Violet Laser
Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
c-Kit, also known as CD117 and stem cell factor receptor, is a 145 kDa transmembrane tyrosine kinase encoded by the c-Kit proto-oncogene. c-Kit acts to regulate a variety of biological responses including cell proliferation, apoptosis, chemotaxis and adhesion. Ligand binding to the extracellular domain leads to autophosphorylation on several tyrosine residues within the cytoplasmic domain, and activation. c-Kit mutations correlate with tumor growth and progression in a variety of cancers including mast cell disease, gastrointestinal stromal tumor, acute myeloid leukemia, Ewing sarcoma, and lung cancer. Phosphorylation at tyrosine 703 of c-Kit allows binding of Grb2 and activation of the Ras-Raf-ERK1 and 2 signaling pathway.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: belly-spot; C-Kit; c-kit proto-oncogene protein; CD117; ckit; dominant spotting; Dominant white spotting; Mast/stem cell growth factor receptor Kit; piebald trait protein; Proto-oncogene c-Kit; proto-oncogene tyrosine-protein kinase Kit; SCFR; soluble KIT variant 1; spotted sterile male; Steel Factor Receptor; Tyrosine-protein kinase Kit; v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene-like protein
Gene Aliases: Bs; c-KIT; CD117; Fdc; Gsfsco1; Gsfsco5; Gsfsow3; KIT; MGF; SCO1; SCO5; Sl; SOW3; Ssm; Tr-kit; W