PTGES3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 10 bp insertion in exon 2 and 1 bp insertion in exon 2.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 10 bp insertion in exon 2 and 1 bp insertion in exon 2
Alternative names
Co chaperone p23, Cytosolic prostaglandin E synthase, Cytosolic prostaglandin E2 synthase, Hsp90 co-chaperone, P23, PTGES 3, Progesterone receptor complex, Progesterone receptor complex p23, Prostaglandin E synthase 3, Prostaglandin E synthase 3 (cytosolic), Sid 3177, TEBP_HUMAN, Telomerase-binding protein p23, Unactive progesterone receptor 23 kD, cPGES
Recommended products
PTGES3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 10 bp insertion in exon 2 and 1 bp insertion in exon 2.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 10 bp insertion in exon 2 and 1 bp insertion in exon 2
- Concentration
Properties
- Gene name
- PTGES3
- 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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The p23 protein also known as prostaglandin E synthase 3 weighs about 18 kDa. It functions as a co-chaperone for the heat shock protein 90 (Hsp90) complex. P23 is found in the cytoplasm and nucleus of cells and shows expression across various tissues including the brain liver and kidney. It plays a role in stabilizing the ATP-bound state of Hsp90 assisting in client protein stabilization and maturation. P23 binding to the Hsp90 complex influences its chaperone activity.
- Biological function summary
P23 assists Hsp90 in protein-folding processes. As part of the Hsp90 chaperone complex p23 contributes to the correct folding and functioning of several essential client proteins. This chaperone activity is important for cellular processes like signal transduction and protein degradation. Without p23 Hsp90's efficiency in maintaining protein homeostasis can decrease highlighting the importance of their interaction in the cell.
- Pathways
The chaperone actions of p23 extensively impact protein signaling pathways and stress response mechanisms. P23 and Hsp90 are important in the Akt/PI3K signaling pathway which regulates cell proliferation and survival. The complex formed by p23 and Hsp90 also involves heat shock proteins like Hsp70 which collaborates in broader cellular stress responses. These connections demonstrate how p23 integrates into cellular regulation and signal transduction networks.
- Associated diseases and disorders
P23's interaction with Hsp90 relates to cancer development and progression. Dysregulation in the p23-Hsp90 interaction can lead to increased stabilization of oncogenic client proteins contributing to tumorigenesis. Additionally p23 implication has been noted in neurodegenerative diseases like Alzheimer's where protein misfolding plays a critical role. Here p23 involvement with proteins like Tau may link its chaperone activity to the pathological protein aggregation seen in such disorders.
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4 product images
Western blot - Human PTGES3 (p23) knockout HEK-293T cell line (ab266791)
Lanes 1-4: Merged signal (red and green). Green - Anti-p23 antibody [EPR3846] ab92503 observed at 23 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.Anti-p23 antibody [EPR3846] ab92503 Anti-p23 antibody [EPR3846] was shown to specifically react with p23 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266791 (knockout cell lysate Human PTGES3 (p23) knockout HEK-293T cell lysate ab258151) was used. Wild-type and p23 knockout samples were subjected to SDS-PAGE. Anti-p23 antibody [EPR3846] ab92503 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-p23 antibody [EPR3846] (Anti-p23 antibody [EPR3846] ab92503) at 1/1000 dilution
Lane 1: Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2: PTGES3 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2: Western blot - Human PTGES3 (p23) knockout HEK-293T cell line (ab266791)
Lane 3: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 4: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
SecondaryAll lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 19 kDa
Observed band size: 23 kDa
Cell Culture - Human PTGES3 (p23) knockout HEK-293T cell line (ab266791)
Representative images PTGES3 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
Sanger Sequencing - Human PTGES3 (p23) knockout HEK-293T cell line (ab266791)
Allele-2: 10 bp insertion in exon 2.
Sanger Sequencing - Human PTGES3 (p23) knockout HEK-293T cell line (ab266791)
Allele-1: 1 bp insertion in exon 2
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