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Anti-Human Vimentin MAb Stable Cell Line-CHO

Cat.No.: CSC-P0215
Cell Line Description: CHO-Anti-Human Vimentin MAb stable cell line is clonally-derived from a CHO cell line, which has been transfected with an anti-human Vimentin 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: Vimentins are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is attached to the nucleus, endoplasmic reticulum, and mitochondria, either laterally or terminally. Involved with LARP6 in the stabilization of type I collagen mRNAs for CO1A1 and CO1A2.
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
Product Molecular Mass: 150 kDa
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 is a human monoclonal antibody for the treatment of brain cancer.
Gene Name: VIM vimentin [ Homo sapiens ]
Official Symbol: VIM
Synonyms: FLJ36605; OTTHUMP00000019224; VIM; Vimentin
Gene ID: 7431
mRNA Refseq: NM_003380.3
Protein Refseq: NP_003371.2
MIM: 193060
UniProt ID: P08670
Chromosome Location: 10p13
Pathway: Alpha6-Beta4 Integrin Signaling Pathway; Apoptosis; Apoptotic cleavage of cellular proteins; Apoptotic execution phase; Aurora B signaling; Caspase cascade in apoptosis; Caspase-mediated cleavage of cytoskeletal proteins;
Function: Identical protein binding; Protein C-terminus binding; Protein binding; Structural constituent of cytoskeleton

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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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