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AB257533

Human MDH2 knockout HEK-293T cell lysate

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MDH2 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 4 bp deletion in exon 4 and Insertion of the selection cassette in exon 4.

View Alternative Names

M MDH, M-OR-1, MDHM_HUMAN, MGC:3559, Malate dehydrogenase, Malate dehydrogenase 2, NAD (mitochondrial), Malate dehydrogenase, mitochondrial, Mitochondrial malate dehydrogenase 2, NAD, mitochondrial

4 Images
Western blot - Human MDH2 knockout HEK-293T cell lysate (AB257533)
  • WB

Lab

Western blot - Human MDH2 knockout HEK-293T cell lysate (AB257533)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : MDH2 knockout HEK293T cell lysate (20 ug)
Lane 3 : K-562 cell lysate (20 ug)
Lane 4 : Human eyeball tissue lysate (20 ug)

ab181873 was shown to specifically react with MDH2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266449 (knockout cell lysate ab257533) was used. Wild-type and MDH2 knockout samples were subjected to SDS-PAGE. ab181873 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-MDH2 antibody [EPR14882(B)] (<a href='/en-us/products/primary-antibodies/mdh2-antibody-epr14882b-ab181873'>ab181873</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

MDH2 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human MDH2 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-mdh2-knockout-hek-293t-cell-line-ab266449'>ab266449</a>)

Lane 3:

K-562 cell lysate at 20 µg

Lane 4:

Human eyeball tissue 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: 36 kDa

Observed band size: 36 kDa

false

Western blot - Human MDH2 knockout HEK-293T cell lysate (AB257533)
  • WB

Lab

Western blot - Human MDH2 knockout HEK-293T cell lysate (AB257533)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : MDH2 knockout HEK293T cell lysate (20 ug)
Lane 3 : K-562 cell lysate (20 ug)
Lane 4 : Human eyeball tissue lysate (20 ug)

ab181857 was shown to specifically react with MDH2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266449 (knockout cell lysate ab257533) was used. Wild-type and MDH2 knockout samples were subjected to SDS-PAGE. ab181857 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-MDH2 antibody [EPR14883(B)] (<a href='/en-us/products/primary-antibodies/mdh2-antibody-epr14883b-ab181857'>ab181857</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

MDH2 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human MDH2 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-mdh2-knockout-hek-293t-cell-line-ab266449'>ab266449</a>)

Lane 3:

K-562 cell lysate at 20 µg

Lane 4:

Human eyeball tissue 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: 36 kDa

Observed band size: 36 kDa

false

Sanger Sequencing - Human MDH2 knockout HEK-293T cell lysate (AB257533)
  • Sanger seq

Unknown

Sanger Sequencing - Human MDH2 knockout HEK-293T cell lysate (AB257533)

Allele-1 : 4 bp deletion in exon 4

Sanger Sequencing - Human MDH2 knockout HEK-293T cell lysate (AB257533)
  • Sanger seq

Unknown

Sanger Sequencing - Human MDH2 knockout HEK-293T cell lysate (AB257533)

Allele-2 : Insertion of the selection cassette 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, 4 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
MDH2
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.

MDH2 also known as malate dehydrogenase 2 or mitochondrial malate dehydrogenase is an enzyme with a molecular mass of approximately 35 kDa. This enzyme catalyzes the conversion of malate to oxaloacetate using NAD+ as a cofactor. It is predominantly expressed in the mitochondria where it plays an important role in cellular respiration. The enzyme enables the malate dehydrogenase reaction which is key for the functioning of the tricarboxylic acid cycle.
Biological function summary

MDH2 participates in the critical process of energy production within the cell. While it does not form a complex itself its activity is intimately connected with other enzymes in mitochondrial energy metabolism. The malate dehydrogenase assay often measures the activity of MDH2 to understand the metabolic status of cells. By facilitating the oxidation of malate MDH2 aids in maintaining the efficiency of the mitochondrial electron transport chain by regenerating NADH.

Pathways

The enzyme is essential in the tricarboxylic acid (TCA) cycle and the malate-aspartate shuttle. In the TCA cycle MDH2 collaborates with enzymes like citrate synthase and isocitrate dehydrogenase to assist in the conversion of acetyl-CoA into energy-rich molecules. The malate-aspartate shuttle on the other hand involves MDH2 working closely with aspartate transaminase to transfer reducing equivalents into the mitochondria. These pathways highlight MDH2’s importance in cellular energy homeostasis.

Mutations or dysregulation in MDH2 have connections to certain metabolic conditions and cancers. For example alterations in MDH2 activity might contribute to conditions like mitochondrial myopathy altering energy metabolism. Moreover MDH2 is associated with NADH-producing enzymes whose dysregulation can support oncogenic pathways in cancer. Understanding these associations helps researchers pursue therapeutic targets that modulate MDH2 activity.
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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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Product promise

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