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

Cat.No.: CSC-P0447
Cell Line Description: CHO-Anti-Human TNF F(ab) stable cell line is clonally-derived from a CHO cell line, which has been transfected with an anti-human TNF 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: Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation.
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: Humanized IgG 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 antibody aliquots should avoid repeated freeze-thaw cycles.
Involvement in Disease: Human monoclonal antibody fragments designed for the treatment of inflammatory diseases.
Gene Name: TNF tumor necrosis factor [ Homo sapiens ]
Official Symbol: TNF
Synonyms: TNF; tumor necrosis factor; DIF; TNFA; TNFSF2; TNF-alpha; TNF superfamily, member 2; APC1 protein; cachectin; TNF, macrophage-derived; TNF-monocyte-derived; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor alpha; tumor necrosis factor-alpha; tumor necrosis factor (TNF superfamily, member 2); Cachectin
Gene ID: 7124
mRNA Refseq: NM_000594
Protein Refseq: NP_000585
MIM: 191160
UniProt ID: P01375
Chromosome Location: 6p21.3
Pathway: Adipocytokine signaling pathway; Adipogenesis; African trypanosomiasis; Allograft rejection; Alzheimer"s disease
Function: cytokine activity; identical protein binding; protease binding; protein binding; transcription regulatory region DNA binding; tumor necrosis factor receptor binding

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C-fusion    N-fusion   Non-tagged
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<1.0 eu/μg    <0.1 eu/μg   <0.01 eu/μg   Not required
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