Human S100P knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human S100P knockout HeLa cell line (AB266026)
ab124743 was shown to react with S100P in wild-type HeLa cells in western blot. Loss of signal was observed when S100P knockout sample was used. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab124743 overnight at 4 °C at a 1 in 1000 dilution. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) secondary antibody at 1/5000 for 1 hour at room temperature before development with Optiblot ECL reagent (ab133456) and imaging.
All lanes:
Western blot - Anti-S100P antibody [EPR6142] (<a href='/en-us/products/primary-antibodies/s100p-antibody-epr6142-ab124743'>ab124743</a>)
Predicted band size: 10 kDa
Observed band size: 9 kDa
false
Exposure time: 20min
- Sanger seq
Unknown
Sanger Sequencing - Human S100P knockout HeLa cell line (AB266026)
Allele-2 : Insertion of the selection cassette in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human S100P knockout HeLa cell line (AB266026)
Allele-1 : 1 bp deletion in exon 1
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
S100P plays a role in intracellular and extracellular calcium signaling. It influences cell survival proliferation and migration. It can function as a monomer but often interacts with other proteins forming complexes such as with RAGE (Receptor for Advanced Glycation End-products) which amplifies signaling cascades that control cellular responses. Its binding to calcium and interaction with RAGE modulate important cellular processes tied to cell cycle and cellular stress response.
Pathways
S100P is closely involved in the MAPK (Mitogen-Activated Protein Kinases) and RAGE signaling pathways. Its interaction with RAGE leads to activation of the MAPK pathway facilitating cellular responses such as growth and survival. S100P also coordinates with other S100 proteins such as S100A4 and S100A9 enhancing its involvement in regulating cellular functions through these pathways. These interactions highlight the importance of S100P in cellular communication and response mechanisms.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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