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Anti-Human EGFR scFv Stable Cell Line-CHO, Clone 3G9

Cat.No.: CSC-P0018
Cell Line Description: CHO-Anti-Human EGFR scFv stable cell line is clonally-derived from a CHO cell line, which has been transfected with an anti-human EGFR scFv gene (Clone 3G9) to allow expression of the scFv. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: Receptor for EGF, but also for other members of the EGF family, as TGF-alpha, amphiregulin, betacellulin, heparin-binding EGF-like growth factor, GP30 and vaccinia virus growth factor. Is involved in the control of cell growth and differentiation. Phosphorylates MUC1 in breast cancer cells and increases the interaction of MUC1 with SRC and CTNNB1/beta-catenin. Isoform 2 may act as an antagonist of EGF action.
Growth Properties: Suspension
Morphology: Epithelial-like
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: Human IgG1 kappa scFv
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 antibody aliquots should avoid repeated freeze-thaw cycles.
Involvement in Disease: This antibody fragments bind to the EGFr Domain III on the cell surface. This locks the receptor in an inactive conformation, making the antibody a inerse agonist. It also is responsible for some antitumor affects through antibody-dependent cellular cytotoxicity (ADCC).
Gene Name: EGFR epidermal growth factor receptor [ Homo sapiens ]
Official Symbol: EGFR
Synonyms: ERBB; HER1; mENA; ERBB1; PIG61; epidermal growth factor receptor; epidermal growth factor receptor; proto-oncogene c-ErbB-1; cell growth inhibiting protein 40; cell proliferation-inducing protein 61; receptor tyrosine-protein kinase erbB-1; avian erythroblastic leukemia viral (v-erb-b) oncogene homolog; EGFR; Cell growth inhibiting protein 40; Cell proliferation inducing protein 61; EGFR antibody; EGFR; EGFR_HUMAN; Epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog); Oncogene ERBB; wa2
Gene ID: 1956
mRNA Refseq: NM_005228.3
Protein Refseq: NP_005219.2
MIM: 131550
UniProt ID: P00533
Chromosome Location: 7p12
Pathway: Adherens junction; Alpha6-Beta4 Integrin Signaling Pathway; Androgen Receptor Signaling Pathway; Arf6 signaling events; Axon guidance; Bladder cancer; Calcium signaling pathway;
Function: ATP binding; MAPK/ERK kinase kinase activity; actin filament binding; double-stranded DNA binding; enzyme binding; epidermal growth factor-activated receptor activity; identical protein binding; contributes_to nitric-oxide synthase regulator activity; protein binding; protein binding; transmembrane signaling receptor activity; transmembrane receptor protein tyrosine kinase activity; signal transducer activity

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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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