S1PR5 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 1 bp deletion in exon 2 and 71 bp deletion in exon 2.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 1 bp deletion in exon 2 and 71 bp deletion in exon 2
Recommended products
S1PR5 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 1 bp deletion in exon 2 and 71 bp deletion in exon 2.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 1 bp deletion in exon 2 and 71 bp deletion in exon 2
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- S1PR5
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Cell culture
- Biosafety level
- EU: 2 US: 2
- Viability
- ~ 80%
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type HeLa cell line (ab271142). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: DMEM (High Glucose) + 10% FBS
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 2x10E4 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 2x10E4 cells/cm2 is recommended.
• Cells should be passaged when they have achieved 80-90% confluence.
• Do not allow the cell density to exceed 7x10E4 cells/cm2.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
EDG8 also known as S1P5 is a receptor with a mass of approximately 41 kDa. It belongs to the G-protein coupled receptor family and specifically binds to sphingosine-1-phosphate (S1P). Researchers often find EDG8 expression in tissues such as the central nervous system spleen and placenta. It plays an important role in mediating cellular responses to extracellular signals through its interaction with ligands.
- Biological function summary
S1P5 functions in cellular growth survival and migration. It is part of a receptor complex that mediates immune response and neurologic functions. This receptor transmits signals that regulate cellular shape and cytoskeletal architecture. S1P5 influences various processes such as lymphocyte trafficking and oligodendrocyte maturation supporting its role in immune and neurological systems.
- Pathways
EDG8 engages in the sphingolipid signaling pathway and is also important in the regulation of the immune system. This receptor interacts with other proteins like G-protein subunits which transduce signals from extracellular factors and promote intracellular responses. The protein S1P5 links to the Akt and MAPK pathways which are important for cell survival and proliferation. These pathways help mediate cellular responses through signal transduction and protein activation.
- Associated diseases and disorders
S1P5 associates notably with multiple sclerosis and some types of cancer. Dysfunction in this receptor's signaling can disrupt normal cell function and immune regulation. S1P5's interactions with proteins like S1P1 and S1P3 further implicate its role in the progression of these diseases. Research continues to explore how modulating EDG8 can offer therapeutic strategies for managing these conditions effectively.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
3 product images
Sanger Sequencing - Human S1PR5 (EDG8) knockout HeLa cell line (ab265613)
Allele-1: 19 bp deletion in exon 2.
Sanger Sequencing - Human S1PR5 (EDG8) knockout HeLa cell line (ab265613)
Allele-2: 1 bp deletion in exon 2.
Sanger Sequencing - Human S1PR5 (EDG8) knockout HeLa cell line (ab265613)
Allele-3: 71 bp deletion in exon 2.
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