Human PTGR1 knockout HeLa cell line
- Advanced Validation
- What is this?
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PTGR1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.
View Alternative Names
15-oxoprostaglandin 13-reductase, FLJ99229, LTB4DH, MGC34943, NADP-dependent leukotriene B4 12-hydroxydehydrogenase, PGR1, PRG-1, PTGR1_HUMAN, Prostaglandin reductase 1, RP11-16L21.1, ZADH3, leukotriene B4 12-hydroxydehydrogenase, zinc binding alcohol dehydrogenase domain containing 3
- WB
Lab
Western blot - Human PTGR1 knockout HeLa cell line (AB265169)
Lanes 1-2 : Merged signal (red and green). Green - ab181131 observed at 38 kDa. Red - loading control ab7291 observed at 50 kDa.
ab181131 Anti-PTGR1 antibody [EPR13451-10] was shown to specifically react with PTGR1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265169 (knockout cell lysate ab258152) was used. Wild-type and PTGR1 knockout samples were subjected to SDS-PAGE. ab181131 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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 (ab216773) and 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-PTGR1 antibody [EPR13451-10] (<a href='/en-us/products/primary-antibodies/ptgr1-antibody-epr13451-10-ab181131'>ab181131</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
PTGR1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human PTGR1 knockout HeLa cell line (ab265169)
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 35 kDa
Observed band size: 38 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PTGR1 knockout HeLa cell line (AB265169)
Allele-2 : Insertion of the selection cassette in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human PTGR1 knockout HeLa cell line (AB265169)
Allele-1 : 2 bp deletion in exon 2.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). 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.
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
PTGR1 participates in metabolizing prostaglandins which play important roles in inflammation and homeostasis. It does not form part of a larger protein complex but works independently to control the levels of active prostaglandins within cells. This activity regulates physiological processes such as inflammation and vascular tone.
Pathways
Prostaglandin metabolism involves PTGR1's function. It integrates into the arachidonic acid pathway. Within this pathway PTGR1 interacts with other enzymes such as cyclooxygenases and lipoxygenases orchestrating the catabolism of prostaglandins and related lipid compounds. This coordination maintains the balance of inflammatory responses.
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
Complementary Products
![Western blot - Anti-PTGR1 antibody [EPR13451-10] (AB181131)](https://content.abcam.com/products/images/ptgr1-antibody-epr13451-10-ab181131--western-blot-img81676.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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