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AB257942

Human ESD knockout HEK-293T cell lysate

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ESD KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 4 and Insertion of the selection cassette in exon 4.

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

4 Images
Western blot - Human ESD knockout HEK-293T cell lysate (AB257942)
  • WB

Lab

Western blot - Human ESD knockout HEK-293T cell lysate (AB257942)

Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2 : ESD knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3 : K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate (20 ug)
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate (20 ug)

ab133631 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 4oC 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 (<a href='/en-us/products/cell-lines/human-esd-knockout-hek-293t-cell-line-ab266360'>ab266360</a>)

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

Western blot - Human ESD knockout HEK-293T cell lysate (AB257942)
  • WB

Lab

Western blot - Human ESD knockout HEK-293T cell lysate (AB257942)

Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2 : ESD knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3 : K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate (20 ug)
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate (20 ug)

ab131211 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 4oC 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 (<a href='/en-us/products/cell-lines/human-esd-knockout-hek-293t-cell-line-ab266360'>ab266360</a>)

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 Sequencing - Human ESD knockout HEK-293T cell lysate (AB257942)
  • Sanger seq

Unknown

Sanger Sequencing - Human ESD knockout HEK-293T cell lysate (AB257942)

Allele-2 : Insertion of the selection cassette in exon 4

Sanger Sequencing - Human ESD knockout HEK-293T cell lysate (AB257942)
  • Sanger seq

Unknown

Sanger Sequencing - Human ESD knockout HEK-293T cell lysate (AB257942)

Allele-1 : 2 bp deletion in exon 4

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

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.

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name
ESD
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Complementary Products

Primary Antibodies

AB69300

Anti-SIAE antibody

Western blot - Anti-SIAE antibody (AB69300)

  • 0
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Primary Antibodies

AB133631

Anti-ESD antibody [EPR8447]

RabMAb|Recombinant|KO Validated

Western blot - Anti-ESD antibody [EPR8447] (AB133631)

  • 0
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Cell Lines & Lysates

AB266360

Human ESD knockout HEK-293T cell line

Advanced Validation

Western blot - Human ESD knockout HEK-293T cell line (AB266360)

  • 0
  • 0
Proteins & Peptides

AB87688

Recombinant Human ESD protein

SDS-PAGE - Recombinant Human ESD protein (AB87688)

  • 0
  • 0

Product promise

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