ESD 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 4 and Insertion of the selection cassette in exon 4.
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, 2 bp deletion in exon 4 and Insertion of the selection cassette in exon 4
Alternative names
EC 3.1.2.12, ESTD_HUMAN, Es-10, Esterase 10, Esterase D, Esterase D formylglutathione hydrolase, FGH, FGH, included, FLJ11763, MGC139609, Methylumbelliferyl acetate deacetylase, OTTHUMP00000018374, OTTHUMP00000040929, S-formylglutathione hydrolase, S-formylglutathione hydrolase, included, Sid 478
Recommended products
ESD 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 4 and Insertion of the selection cassette in exon 4.
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, 2 bp deletion in exon 4 and Insertion of the selection cassette in exon 4
- Concentration
Properties
- Gene name
- ESD
- 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
ESD also known as esterase D or S-formylglutathione hydrolase is a protein with a mass of approximately 31 kDa. ESD is expressed in various tissues and cells including HEK-293T cells. Cell lysate studies often utilize the ESD line to analyze its enzymatic activity and expression profile. The protein plays a mechanical role in hydrolyzing esters of glutathione contributing to the detoxification processes within the cell.
- Biological function summary
ESD contributes to maintaining cellular homeostasis through its role in detoxification and response to oxidative stress. It interacts with the degradation of S-formylglutathione a byproduct formed during alcohol metabolism. ESD is not typically associated with a larger protein complex but its enzymatic function is important for cellular health and efficient elimination of toxic metabolites protecting cells from damage and supporting metabolic pathways.
- Pathways
ESD's enzymatic activity is integrated into the glutathione metabolism and alcohol metabolism pathways. Here ESD acts alongside enzymes like glutathione S-transferase to convert harmful aldehydes into less toxic forms aiding in cellular resilience. The protein helps modulate oxidative stress responses cooperating indirectly with other detoxification proteins within these pathways to maintain optimal cellular function.
- Associated diseases and disorders
ESD associations include certain forms of cancer and neurodegenerative diseases. Abnormal ESD expression or activity can result in disturbed detoxification leading to oxidative stress and cell damage. In cancer ESD has been implicated in tumor progression potentially linked with other proteins like GSTP1 which also participates in detoxification. In neurodegenerative conditions defective detoxification pathways involving ESD may contribute to neuronal injury and disease progression making it a potential target for therapeutic exploration.
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5 product images
Western blot - Human ESD knockout HEK-293T cell line (ab266360)
Lanes 1-4: Merged signal (red and green). Green - Anti-ESD antibody [EPR8446] ab131211 observed at 31 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.Anti-ESD antibody [EPR8446] ab131211 Anti-ESD antibody [EPR8446] was shown to specifically react with ESD in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266360 (knockout cell lysate Human ESD knockout HEK-293T cell lysate ab257942) was used. Wild-type and ESD knockout samples were subjected to SDS-PAGE. Anti-ESD antibody [EPR8446] ab131211 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C 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-ESD antibody [EPR8446] (Anti-ESD antibody [EPR8446] ab131211) 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: ESD 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 ESD knockout HEK-293T cell line (ab266360)
Lane 3: K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate at 20 µg
Lane 4: Jurkat (Human T cell leukemia cell line from peripheral blood) 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: 31 kDa
Observed band size: 31 kDa
Western blot - Human ESD knockout HEK-293T cell line (ab266360)
Lanes 1-4: Merged signal (red and green). Green - Anti-ESD antibody [EPR8447] ab133631 observed at 31 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.Anti-ESD antibody [EPR8447] ab133631 Anti-ESD antibody [EPR8447] was shown to specifically react with ESD in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266360 (knockout cell lysate Human ESD knockout HEK-293T cell lysate ab257942) was used. Wild-type and ESD knockout samples were subjected to SDS-PAGE. Anti-ESD antibody [EPR8447] ab133631 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C 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-ESD antibody [EPR8447] (Anti-ESD antibody [EPR8447] ab133631) 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: ESD 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 ESD knockout HEK-293T cell line (ab266360)
Lane 3: K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate at 20 µg
Lane 4: Jurkat (Human T cell leukemia cell line from peripheral blood) 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: 31 kDa
Observed band size: 31 kDa
Cell Culture - Human ESD knockout HEK-293T cell line (ab266360)
Representative images ESD 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 ESD knockout HEK-293T cell line (ab266360)
Allele-1: 2 bp deletion in exon 4
Sanger Sequencing - Human ESD knockout HEK-293T cell line (ab266360)
Allele-2: Insertion of the selection cassette in exon 4.
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