|Anti-Human EGFR Stable Cell Line
- Cell Line Description
- Receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses. Known ligands include EGF, TGFA/TGF-alpha, amphiregulin, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF. Ligand binding triggers receptor homo- and/or heterodimerization and autophosphorylation on key cytoplasmic residues. The phosphorylated receptor recruits adapter proteins like GRB2 which in turn activates complex downstream signaling cascades. Activates at least 4 major downstream signaling cascades including the RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCgamma-PKC and STATs modules. May also activate the NF-kappa-B signaling cascade. Also directly phosphorylates other proteins like RGS16, activating its GTPase activity and probably coupling the EGF receptor signaling to the G protein-coupled receptor signaling. Also phosphorylates MUC1 and increases its interaction with SRC and CTNNB1/beta-catenin.
- Growth Properties
- Complete growth medium: Serum-free Medium
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
- 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 15ml conical tube containing 10 ml pre-warmed media.
4. Centrifuge for 3 minutes 1000 xg to pellet cells and remove the supernatant.
5. Add 14 ml of media and transfer cells to a T25 flask or a 100 mm culture dish.
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 re-attach to the surface over a period of several days in culture at 37°C.
- 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 Status: Negative (MycoAlert Kit)
- Freeze Medium
- Complete growth medium 90%; DMSO, 10%
- 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.
- Product Purity
- >98.0% as determined by Analysis by RP-HPLC & analysis by SDS-PAGE.
- Product Storage
- It should be stored in liquid or vapor phase nitrogen. Reconstituted antibody aliquots should avoid repeated freeze-thaw cycles.
- Involvement in Disease
- Matuzumab is a humanized monoclonal antibody for the treatment of cancer.
- EGFR; epidermal growth factor receptor; epidermal growth factor receptor (avian erythroblastic leukemia viral (v erb b) oncogene homolog) , ERBB; ERBB1; erythroblastic leukemia viral (v erb b) oncogene homolog (avian); 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; ERBB; HER1; mENA; PIG61;
- Adherens junction, organism-specific biosystem; Adherens junction, conserved biosystem; Alpha6-Beta4 Integrin Signaling Pathway, organism-specific biosystem; Androgen Receptor Signaling Pathway, organism-specific biosystem; Arf6 signaling events, organism-specific biosystem; Axon guidance, organism-specific biosystem; Bladder cancer, organism-specific biosystem;
- ATP binding; MAPK/ERK kinase kinase activity; actin filament binding; double-stranded DNA binding; enzyme binding; epidermal growth factor-activated receptor activity; epidermal growth factor-activated receptor activity; identical protein binding; contributes_to nitric-oxide synthase regulator activity; nucleotide binding; protein binding; protein heterodimerization activity; protein phosphatase binding; protein tyrosine kinase activity; protein tyrosine kinase activity; protein tyrosine kinase activity; receptor activity; receptor signaling protein tyrosine kinase activity; signal transducer activity; transmembrane receptor protein tyrosine kinase activity; transmembrane receptor protein tyrosine kinase activity; transmembrane signaling receptor activity;
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