Human ESD knockout HEK-293T cell line
- Advanced Validation
- What is this?
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ESD KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 4 and Insertion of the selection cassette in exon 4. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View 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
- WB
Lab
Western blot - Human ESD knockout HEK-293T cell line (AB266360)
Lanes 1-4 : Merged signal (red and green). Green - ab133631 observed at 31 kDa. Red - loading control ab8245 observed at 36 kDa.
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 ab257942) was used. Wild-type and ESD knockout samples were subjected to SDS-PAGE. ab133631 and 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 (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-ESD antibody [EPR8447] (<a href='/en-us/products/primary-antibodies/esd-antibody-epr8447-ab133631'>ab133631</a>) 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
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: 31 kDa
Observed band size: 31 kDa
false
- WB
Lab
Western blot - Human ESD knockout HEK-293T cell line (AB266360)
Lanes 1-4 : Merged signal (red and green). Green - ab131211 observed at 31 kDa. Red - loading control ab8245 observed at 36 kDa.
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 ab257942) was used. Wild-type and ESD knockout samples were subjected to SDS-PAGE. ab131211 and 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 (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-ESD antibody [EPR8446] (<a href='/en-us/products/primary-antibodies/esd-antibody-epr8446-ab131211'>ab131211</a>) 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
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: 31 kDa
Observed band size: 31 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ESD knockout HEK-293T cell line (AB266360)
Allele-2 : Insertion of the selection cassette in exon 4.
- Sanger seq
Unknown
Sanger Sequencing - Human ESD knockout HEK-293T cell line (AB266360)
Allele-1 : 2 bp deletion in exon 4
- Cell Culture
Lab
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.
Reactivity data
Product details
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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com