KIT antibody

Catalog No. ABIN114259

This Rat Monoclonal antibody specifically detects KIT in FACS, IF and IHC (fro). It exhibits reactivity toward Mouse.

Quick Overview for KIT antibody (ABIN114259)

Target: KIT (Mast/stem Cell Growth Factor Receptor (KIT))

Reactivity: Mouse

Host: Rat

Clonality: Monoclonal

Conjugate: This KIT antibody is un-conjugated

Application: Flow Cytometry (FACS), Immunofluorescence (IF), Immunohistochemistry (Frozen Sections) (IHC (fro))

Clone: ACK4

Product Details anti-KIT Antibody

Specificity: This monoclonal antibody recognizes the receptor tyrosine kinase, c-kit. The ligand for this receptor is steel factor (stem cell factor), which exists in both soluble and membrane form. The interaction between steel factor and c-kit is essential for the development of hematopoietic, gonadal and pigment stem cells. c-kit positive cells are a subset of CD34+ hematopoietic precursor cells and it is expressed on 5-10 % of total adult bone marrow cells.

Purification: Protein G Chromatography.

Immunogen: IL-3 dependent mast cells derived from WB- +/+ mice. Donor: Wistar spleen. Fusion Partner: X63.653. Ag8.

Isotype: IgG2a

Application Details

Application Notes: Flow Cytometry. (Reported to be useful in immunohistochemistry on frozen and polyester wax sections. )
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation . 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of thissuspension to each tube (each tube will then contain 1x10e6 cells, representing 1 test). 4. To each tube, add 0. 2 - 0. 1 µg of this AB per 10e6 cells. 5. Vortex the tubes to ensure thorough mixing of antibody and cells. 6. Incubate the tubes for 30 minutes at 4°C. 7. Wash 2 times at 4°C. 8. Add 100 µl of secondary antibody (PE Goat anti-rat IgG (H+L)) at appropriate dilution. 9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes areprotected from light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C in media B. 11. Resuspend the cell pellet in 50 µl ice cold media B. 12. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidiumiodide at 0. 5 mg/ml in PBS. This stains dead cells by intercalating in DNA. Media: A. Phosphate buffered saline (pH 7. 2) + 5% normal serum of host species + sodium azide(100 µl of 2M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7. 2) + 0. 5% Bovine serum albumin + sodium azide (100µl of 2M sodium azide in 100 mls). Results - Tissue Distribution: Mouse Strain: BALB/cCell Concentration: 1x10e6 cells per testAntibody Concentration Used: 0. 2 µg/10e6 cellsIsotypic Control: Purified Rat IgG2aResults - Strain Distribution: Cell Concentration: 1x10e6 cells per testAntibody Concentration Used: 0. 2 µg /10e6 cellsStrains Tested: AKR, BALB/c, C3H/He, C57BL/6Positive: AKR, BALB/c, C3H/He, C57BL/6Negative: none

Restrictions: For Research Use only

Handling

Concentration: 1.0 mg/mL

Buffer: PBS containing 0.02 % Sodium Azide as preservative.

Preservative: Sodium azide

Precaution of Use: This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freezing and thawing.

Storage: 4 °C/-20 °C

Storage Comment: Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.

Target Details for KIT

Target: KIT (Mast/stem Cell Growth Factor Receptor (KIT))

Alternative Name: CD117 / C-Kit

Background: C-Kit is a transmembrane tyrosine kinase encoded by the cKit 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. Mutations in c-Kit have been found to be important for tumor growth and progression in a variety of cancers including mast cell diseases, gastrointestinal stromal tumor, acute myeloid leukemia, Ewing sarcoma and lung cancer. Phosphorylation at tyrosine 721 of c-Kit allows binding and activation of PI3 kinase.Synonyms: KIT, Mast/stem cell growth factor receptor, Proto-Oncogene Tyrosine-Protein Kinase, SCFR

Gene ID: 16590

NCBI Accession: NP_066922

UniProt: P05532

Pathways: RTK Signaling, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Sensory Perception of Sound, Stem Cell Maintenance, Production of Molecular Mediator of Immune Response, Regulation of long-term Neuronal Synaptic Plasticity