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Anti-Human INSR MAb Stable Cell Line-CHO

Cat.No.: CSC-P0875
Cell Line Description: CHO-Anti-Human INSR MAb stable cell line is clonally-derived from a CHO cell line, which has been transfected with an Anti-human INSR MAb gene to allow expression of the MAb. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: Insulin receptor mediates the biological activities of insulin by regulating multiple signaling pathways through activation of a series of phosphorylation cascades. The human insulin receptor is a heterotetrameric membrane glycoprotein consisting of disulfide-linked subunits in a ?-a-a-? configuration. The ?-subunit (95kDa) possesses a single transmembrane domain with tyrosine kinase acivity, whereas the a-subunit (135kDa) is completely extracellular. The alpha subunits each contain insulin binding sites and are entirely extracellular in localization. The beta subunits each possess an extracellular domain, a single transmembrane domain, and a cytoplasmic tyrosine kinase domain. Binding of insulin to the alpha subunits induces a conformation change in the receptor which activates the kinase domain, stimulating tyrosine autophosphorylation of the receptor and tyrosine phosphorylation of at least five different insulin receptor substrates designated IRS-1-4, and Shc.
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: IgG
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 binds to INSR.
Gene Name: INSR insulin receptor [ Homo sapiens ]
Official Symbol: INSR
Synonyms: INSR; insulin receptor; CD220; IR; HHF5;
mRNA Refseq: NM_000208
Protein Refseq: NP_000199
MIM: 147670
UniProt ID: P06213
Chromosome Location: 19p13.3-p13.2
Pathway: Adherens junction, organism-specific biosystem; Adherens junction, conserved biosystem; Aldosterone-regulated sodium reabsorption, organism-specific biosystem; Aldosterone-regulated sodium reabsorption, conserved biosystem; IRS activation, organism-specific biosystem; IRS-mediated signalling, organism-specific biosystem; IRS-related events, organism-specific biosystem
Function: 3-phosphoinositide-dependent protein kinase binding; ATP binding; GTP binding; PTB domain binding; SH2 domain binding; insulin binding; insulin binding; insulin binding; insulin receptor substrate binding; insulin-activated receptor activity; insulin-like

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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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