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Anti-Human CD3 epsilon scFv Stable Cell Line-CHO

Cat.No.: CSC-P0421
Cell Line Description: CHO-Anti-Human CD3 epsilon scFv stable cell line is clonally-derived from a CHO cell line, which has been transfected with an anti-human CD3 epsilon scFv gene 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: The protein encoded by this gene is the CD3-epsilon polypeptide, which together with CD3-gamma, -delta and -zeta, and the T-cell receptor alpha/beta and gamma/delta heterodimers, forms the T-cell receptor-CD3 complex. This complex plays an important role in coupling antigen recognition to several intracellular signal-transduction pathways. The genes encoding the epsilon, gamma and delta polypeptides are located in the same cluster on chromosome 11. The epsilon polypeptide plays an essential role in T-cell development. Defects in this gene cause immunodeficiency. This gene has also been linked to a susceptibility to type I diabetes in women.
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 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.
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 95%; DMSO, 5%
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 Storage: It should be stored at –20 °C. Reconstituted antibody aliquots should avoid repeated freeze-thaw cycles.
Involvement in Disease: Monoclonal antibody fragments binds to CD3 epsilon.
Gene Name: CD3E CD3e molecule, epsilon (CD3-TCR complex) [ Homo sapiens ]
Official Symbol: CD3E
Synonyms: CD3e molecule, epsilon (CD3-TCR complex); TCRE; CD3e antigen, epsilon polypeptide (TiT3 complex); T-cell antigen receptor complex, epsilon subunit of T3; T3E; T-cell surface glycoprotein CD3 epsilon chain; T-cell surface antigen T3/Leu-4 epsilon chain; CD3e antigen; CD3-epsilon; CD3E
Gene ID: 916
mRNA Refseq: NM_000733
Protein Refseq: NP_000724
MIM: 186830
UniProt ID: P07766
Chromosome Location: 11q23
Pathway: Adaptive Immune System; CXCR4-mediated signaling events; Chagas disease (American trypanosomiasis); Costimulation by the CD28 family; Downstream TCR signaling; Downstream signaling in naive CD8+ T cells; Generation of second messenger molecules;
Function: SH3 domain binding; T cell receptor binding; protein heterodimerization activity; protein kinase binding; receptor signaling complex scaffold activity; receptor signaling protein activity; transmembrane signaling receptor 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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