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Anti-Human Selectin P scFv Stable Cell Line-CHO

Cat.No.: CSC-P0424
Cell Line Description: CHO-Anti-Human selectin P scFv stable cell line is clonally-derived from a CHO cell line, which has been transfected with an anti-human selectin P 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: Ca(2+)-dependent receptor for myeloid cells that binds to carbohydrates on neutrophils and monocytes. Mediates the interaction of activated endothelial cells or platelets with leukocytes. The ligand recognized is sialyl-Lewis X. Mediates rapid rolling of leukocyte rolling over vascular surfaces during the initial steps in inflammation through interaction with PSGL1.
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 IgG4 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: Humanized monoclonal antibody fragments that binds to selectin P.
Gene Name: SELP selectin P (granule membrane protein 140kDa, antigen CD62) [ Homo sapiens ]
Official Symbol: SELP
Synonyms: selectin P (granule membrane protein 140kDa, antigen CD62); GMP-140; GRMP; LECAM3; PADGEM; CD62 antigen-like family member P; CD62; selectin P (granule membrane protein 140kD, antigen CD62); CD62P; P-selectin; GMP140; granulocyte membrane protein; PSEL; platelet alpha-granule membrane protein; Granule membrane protein 140; GMRP; Leukocyte-endothelial cell adhesion molecule 3; CD62P antigen; Platelet activation dependent granule-external membrane protein; SELP
Gene ID: 6403
mRNA Refseq: NM_003005
Protein Refseq: NP_002996
MIM: 173610
UniProt ID: P16109
Chromosome Location: 11q22-q25
Pathway: Cell adhesion molecules (CAMs); Cell surface interactions at the vascular wall; Hemostasis; IL-3 Signaling Pathway; IL4-mediated signaling events; Malaria; Platelet activation, signaling and aggregation;
Function: calcium-dependent protein binding; eukaryotic cell surface binding; fucose binding; glycoprotein binding; glycosphingolipid binding; heparin binding; lipopolysaccharide binding; oligosaccharide binding; protein binding; sialic acid binding

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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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