|Anti-Human Von Willebrand Factor Stable Cell Line
- Cell Line Description
- Important in the maintenance of hemostasis, it promotes adhesion of platelets to the sites of vascular injury by forming a molecular bridge between sub-endothelial collagen matrix and platelet-surface receptor complex GPIb-IX-V. Also acts as a chaperone for coagulation factor VIII, delivering it to the site of injury, stabilizing its heterodimeric structure and protecting it from premature clearance from plasma.
- 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
- Caplacizumab (INN) is a humanized monoclonal that binds to VWF.
- VWF; von Willebrand factor; F8VWF; coagulation factor VIII VWF; VWD;
- Chromosome Location
- Blood Clotting Cascade, organism-specific biosystem; Complement and Coagulation Cascades, organism-specific biosystem; Complement and coagulation cascades, organism-specific biosystem; Complement and coagulation cascades, conserved biosystem; ECM-receptor interaction, organism-specific biosystem; ECM-receptor interaction, conserved biosystem; Focal Adhesion, organism-specific biosystem;
- chaperone binding; collagen binding; glycoprotein binding; immunoglobulin binding; integrin binding; protease binding; protease binding; protein N-terminus binding; protein binding; protein homodimerization activity;
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