VDR KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 3 and 2 bp deletion in exon 3.
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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, 1 bp insertion in exon 3 and 2 bp deletion in exon 3
Alternative names
1 25 dihydroxyvitamin D3 receptor, 25-dihydroxyvitamin D3 receptor, Member 1, NR1I1, Nuclear receptor subfamily 1 group I member 1, PPP1R163, Protein phosphatase 1, regulatory subunit 163, VDR_HUMAN, Vitamin D (1,25- dihydroxyvitamin D3) receptor, Vitamin D hormone receptor, Vitamin D nuclear receptor variant 1, Vitamin D receptor, Vitamin D3 receptor
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VDR KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 3 and 2 bp deletion in exon 3.
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, 1 bp insertion in exon 3 and 2 bp deletion in exon 3
- Antibiotic resistance
- Puromycin 1µg/mL
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- VDR
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- 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
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.
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
Western blot - Human VDR (Vitamin D Receptor) knockout HeLa cell line (ab265430)
Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade ab109234 Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade was shown to specifically react with Vitamin D Receptor in HeLa wild-type cells (ab255928). Loss of signal was observed when knockout cell line ab265430 (knockout cell lysate Human VDR (Vitamin D Receptor) knockout HeLa cell lysate ab257796) was used. Wild-type and Vitamin D Receptor knockout samples were subjected to SDS-PAGE. Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade ab109234 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade (Anti-Vitamin D Receptor antibody [EPR4552] - ChIP Grade ab109234) at 1/1000 dilution
Lane 1: Wild-type HeLa lysate at 20 µg
Lane 2: Vitamin D Receptor knockout HeLa lysate at 20 µg
Lane 2: Western blot - Human VDR (Vitamin D Receptor) knockout HeLa cell line (ab265430)
Lane 3: U-937 lysate at 20 µg
Lane 4: SH-SY5Y lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 48 kDa
Sanger Sequencing - Human VDR (Vitamin D Receptor) knockout HeLa cell line (ab265430)
Allele-2: 1 bp insertion in exon 3.
Sanger Sequencing - Human VDR (Vitamin D Receptor) knockout HeLa cell line (ab265430)
Allele-1: 2 bp deletion in exon 3.
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