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Anti-Human KIT F(ab) Stable Cell Line-CHO

Cat.No.: CSC-P0660
Cell Line Description: CHO-Anti-Human KIT F(ab) stable cell line is clonally-derived from a CHO cell line, which has been transfected with an Anti-human KIT F(ab) gene to allow expression of the F(ab). It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: Tyrosine-protein kinase that acts as cell-surface receptor for the cytokine KITLG/SCF and plays an essential role in the regulation of cell survival and proliferation, hematopoiesis, stem cell maintenance, gametogenesis, mast cell development, migration and function, and in melanogenesis. In response to KITLG/SCF binding, KIT can activate several signaling pathways. Phosphorylates PIK3R1, PLCG1, SH2B2/APS and CBL. Activates the AKT1 signaling pathway by phosphorylation of PIK3R1, the regulatory subunit of phosphatidylinositol 3-kinase. Activated KIT also transmits signals via GRB2 and activation of RAS, RAF1 and the MAP kinases MAPK1/ERK2 and/or MAPK3/ERK1. Promotes activation of STAT family members STAT1, STAT3, STAT5A and STAT5B. Activation of PLCG1 leads to the production of the cellular signaling molecules diacylglycerol and inositol 1,4,5-trisphosphate. KIT signaling is modulated by protein phosphatases, and by rapid internalization and degradation of the receptor. Activated KIT promotes phosphorylation of the protein phosphatases PTPN6/SHP-1 and PTPRU, and of the transcription factors STAT1, STAT3, STAT5A and STAT5B. Promotes phosphorylation of PIK3R1, CBL, CRK (isoform Crk-II), LYN, MAPK1/ERK2 and/or MAPK3/ERK1, PLCG1, SRC and SHC1.
Growth Properties: Suspension
Morphology: Epithelial-like
Species: Human
Host Cell: CHO
Propagation: Complete growth medium: Serum-free MediumAtmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37 °C
Starting Cells From Frozen Cell Stock: 1. Remove the packaging cell lines from liquid nitrogen and carry out a quick thaw. Float the cells in the 37 °C water bath for 2 minutes until nearly (80%) thawed. Once cells are thawed, it is important to dilute the cells 1:10 in growth media immediately to reduce the potentially toxic effects of the DMSO preservative on the cells.2. Clean the outside of the vial with 70% ethanol before opening.3. Remove the cells from the vial and add slowly into a 15 ml conical tube containing 10 ml pre-warmed media. 4. Centrifuge for 5 minutes 300 xg to pellet cells and remove the supernatant.5. Add 14 ml of media and transfer cells to a T25 shaker flask.6. Place the cells in the 37 °C incubator with 5% CO2.7. Allow incubation for 3-4 days to reach confluence. The cells will grow well over a period of several days in culture at 37 °C.
Subculturing: 1. Centrifuge for 5 minutes 300 xg and discard culture medium.2. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.3. Add appropriate aliquots of the cell suspension to new culture vessels.Incubate cultures at 37 °C.Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended Medium Renewal: 2 to 3 times per week
Mycoplasma: Mycoplasma Status: Negative (MycoAlert Kit)
Freeze Medium: Complete growth medium 90%; DMSO, 10%
Storage: Liquid nitrogen vapor phase
Preservation: 1. Detach cells from culture dish according to the Sub-Culture Procedure.2. Resuspend cells at a density of 5 x 10^6 cells/mL in freeze medium.3. Aliquot 1 mL cells into cryogenic vials.4. Place vials in a freezing container and store at –80 °C overnight.5. Transfer vials to liquid nitrogen for long term storage. If properly stored, cells should remain stable for years.
Safety Considerations: The following safety precautions should be observed.1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum.2. No eating, drinking or smoking while handling the stable line.3. Wash hands after handling the stable line and before leaving the lab.4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.5. All waste should be considered hazardous.6. Dispose of all liquid waste after each experiment and treat with bleach.
Ship: Dry ice
Product Type: Fab
Product Purity: HPLC >98%, One strip for SDS-PAGE.
Product Activity/Specificity: Tested positive against native human antigen.
Product Storage: It should be stored at –20 °C. Reconstituted protein aliquots should avoid repeated freeze-thaw cycles.
Involvement in Disease: Monoclonal antibody fragments binds to KIT.
Gene Name: KIT v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog [ Homo sapiens ]
Official Symbol: KIT
Synonyms: KIT; v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog; PBT, piebald trait; mast/stem cell growth factor receptor Kit; C Kit; CD117; SCFR; p145 c-kit; proto-oncogene c-Kit; piebald trait protein; soluble KIT variant 1; tyrosine-protein kinase Kit; proto-oncogene tyrosine-protein kinase Kit; v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene-like protein; PBT; C-Kit;
Gene ID: 3815
mRNA Refseq: NM_000222
Protein Refseq: NP_000213
MIM: 164920
UniProt ID: P10721
Chromosome Location: 4q11-q12
Pathway: Acute myeloid leukemia, organism-specific biosystem; Acute myeloid leukemia, conserved biosystem; C-MYB transcription factor network, organism-specific biosystem; Cytokine-cytokine receptor interaction, organism-specific biosystem; Cytokine-cytokine receptor interaction, conserved biosystem; Endocytosis, organism-specific biosystem; Endocytosis, conserved biosystem
Function: ATP binding; cytokine bindingetal ion binding; nucleotide binding; protease binding; protein binding; protein homodimerization activity; protein tyrosine kinase activity; receptor activity; receptor signaling protein tyrosine kinase activity; stem cell fa

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Optional requirements on this protein    +Expand
C-fusion    N-fusion   Non-tagged
His    GST   Fc   Others
<1.0 eu/μg    <0.1 eu/μg   <0.01 eu/μg   Not required
Monomer Isolation    Dimer Isolation    Not required
>80% by SDS-PAGE    >90% by SDS-PAGE   >95% by SDS-PAGE   Others

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