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AB257396

Human COMT knockout HEK-293T cell lysate

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COMT KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 3.

View Alternative Names

COMT_HUMAN, Catechol O-methyltransferase, EC 2.1.1.6

3 Images
Western blot - Human COMT knockout HEK-293T cell lysate (AB257396)
  • WB

Lab

Western blot - Human COMT knockout HEK-293T cell lysate (AB257396)

Lane 1 : Wild-type HEK-293T cell lysate (20 μg)

Lane 2 : COMT knockout HEK-293T cell lysate (20 μg)

Lanes 1-2 : Merged signal (red and green). Green - ab124813 observed at 28 kDa. Red - loading control ab8245 observed at 37 kDa.

ab124813 Anti-COMT antibody [EPR6491(B)] was shown to specifically react with COMT in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266537 (knockout cell lysate ab257396) was used. Wild-type and COMT knockout samples were subjected to SDS-PAGE. ab124813 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-COMT antibody [EPR6491(B)] (<a href='/en-us/products/primary-antibodies/comt-antibody-epr6491b-ab124813'>ab124813</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

COMT knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human COMT knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-comt-knockout-hek-293t-cell-line-ab266537'>ab266537</a>)

Predicted band size: 30 kDa

Observed band size: 28 kDa

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Western blot - Human COMT knockout HEK-293T cell lysate (AB257396)
  • WB

Lab

Western blot - Human COMT knockout HEK-293T cell lysate (AB257396)

Lane 1 : Wild-type HEK-293T cell lysate (20 μg)

Lane 2 : COMT knockout HEK-293T cell lysate (20 μg)

Lanes 1-2 : Merged signal (red and green). Green - ab126618 observed at 24-28 kDa. Red - loading control ab8245 observed at 37 kDa.

ab126618 Anti-COMT antibody [EPR6490] was shown to specifically react with COMT in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266537 (knockout cell lysate ab257396) was used. Wild-type and COMT knockout samples were subjected to SDS-PAGE. ab126618 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-COMT antibody [EPR6490] (<a href='/en-us/products/primary-antibodies/comt-antibody-epr6490-ab126618'>ab126618</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

COMT knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human COMT knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-comt-knockout-hek-293t-cell-line-ab266537'>ab266537</a>)

Predicted band size: 30 kDa

Observed band size: 24-28 kDa

false

Sanger Sequencing - Human COMT knockout HEK-293T cell lysate (AB257396)
  • Sanger seq

Unknown

Sanger Sequencing - Human COMT knockout HEK-293T cell lysate (AB257396)

Homozygous : 1 bp insertion in exon 3

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, Homozygous: 1 bp insertion in exon 3.

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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
COMT
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
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.

COMT also known as catechol-O-methyltransferase is an enzyme responsible for the methylation of catechol compounds including neurotransmitters like dopamine epinephrine and norepinephrine. This reaction requires S-adenosylmethionine as a methyl donor. COMT primarily has two forms: a soluble form and a membrane-bound form with masses around 24 kDa and 30 kDa respectively. This protein shows significant expression in the liver brain and kidneys. COMT expression is also noted in cell lines such as HEK293T where it plays an important role in catecholamine metabolism.
Biological function summary

Catechol-O-methyltransferase serves as a regulator of neurotransmitter levels in the central nervous system and peripheral tissues. It does not appear to form part of a larger protein complex functioning mainly as a singular entity for metabolizing catechols. Through its action COMT impacts processes such as mood regulation cognition and stress response. Its role in degrading dopamine is especially significant in regions like the prefrontal cortex where dopamine plays an important role in executive functions.

Pathways

COMT participates in the catecholamine degradation pathway and impacts dopaminergic signaling. It works alongside enzymes such as monoamine oxidase (MAO) to regulate levels of neurotransmitters. COMT's activity influences the synaptic presence of dopamine and its interaction in signaling pathways. Moreover it indirectly affects the balance within the dopaminergic system which is important for normal neurological function and behavior.

COMT has links to psychiatric and neurological conditions such as schizophrenia and Parkinson’s disease. Variations in COMT function can affect dopamine metabolism which may contribute to the etiology of these disorders. Additionally proteins like MAO and dopamine transporter (DAT) also connect to COMT through their shared involvement in neurotransmitter regulation. Alterations in COMT activity can influence disease severity and patient response to treatment underlining its importance in understanding these disorders.
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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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