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AB261657

Human SDHA knockout HEK-293 cell lysate

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SDHA KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 99%.

View Alternative Names

CMD1GG, DHSA_HUMAN, FP, Flavoprotein subunit of complex II, PGL5, SDH 1, SDH 2, SDHF, Succinate dehydrogenase [ubiquinone] flavoprotein subunit, Succinate dehydrogenase [ubiquinone] flavoprotein subunit mitochondrial, Succinate dehydrogenase complex flavoprotein subunit, Succinate dehydrogenase complex flavoprotein subunit A, Succinate dehydrogenase complex flavoprotein subunit precursor, Succinate dehydrogenase complex subunit A, Succinate dehydrogenase complex subunit A flavoprotein, Succinate dehydrogenase complex subunit A flavoprotein (Fp)

4 Images
Western blot - Human SDHA knockout HEK-293 cell lysate (AB261657)
  • WB

Supplier Data

Western blot - Human SDHA knockout HEK-293 cell lysate (AB261657)

Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate 20 ug

Lane 2 : SDHA knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate 20 ug

Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 20 ug

Lane 4 : Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate 20 ug

Lanes 1 - 4 : Merged signal (red and green). Green - ab137040 observed at 72 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab137040 was shown to specifically react with SDHA in wild-type HEK-293 cells as signal was lost in SDHA knockout cell line ab261853 (knockout cell lysate ab261657). Wild-type and SDHA knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab137040 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-SDHA antibody [EPR9043(B)] (<a href='/en-us/products/primary-antibodies/sdha-antibody-epr9043b-ab137040'>ab137040</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

SDHA knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

Western blot - Human SDHA knockout HEK-293 cell line (<a href='/en-us/products/cell-lines/human-sdha-knockout-hek-293-cell-line-ab261853'>ab261853</a>)

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 72 kDa

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Western blot - Human SDHA knockout HEK-293 cell lysate (AB261657)
  • WB

Supplier Data

Western blot - Human SDHA knockout HEK-293 cell lysate (AB261657)

Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate 20 ug

Lane 2 : SDHA knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate 20 ug

Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 20 ug

Lane 4 : Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate 20 ug

Lanes 1 - 4 : Merged signal (red and green). Green - ab139181 observed at 72 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab139181 was shown to recognize SDHA in wild-type HEK-293 cells as signal was lost at the expected MW in SDHA knockout cell line ab261853 (knockout cell lysate ab261657). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and SDHA knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab139181 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-SDHA antibody [EPR9042(B)] (<a href='/en-us/products/primary-antibodies/sdha-antibody-epr9042b-ab139181'>ab139181</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

SDHA knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

Western blot - Human SDHA knockout HEK-293 cell line (<a href='/en-us/products/cell-lines/human-sdha-knockout-hek-293-cell-line-ab261853'>ab261853</a>)

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 72 kDa

Observed band size: 72 kDa

false

Western blot - Human SDHA knockout HEK-293 cell lysate (AB261657)
  • WB

Lab

Western blot - Human SDHA knockout HEK-293 cell lysate (AB261657)

Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate 20 ug

Lane 2 : SDHA knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate 20 ug

Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 20 ug

Lane 4 : Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate 20 ug

ab198493 was shown to specifically react with SDHA in wild-type HEK-293 cells as signal was lost in SDHA knockout cell line ab261853 (knockout cell lysate ab261657). Wild-type and SDHA knockout samples were subjected to SDS-PAGE. ab198493 and ab181602 (Rabbit monoclonal to GAPDH - Loading Control loading) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

All lanes:

Western blot - HRP Anti-SDHA antibody [2E3GC12FB2AE2] (<a href='/en-us/products/primary-antibodies/hrp-sdha-antibody-2e3gc12fb2ae2-ab198493'>ab198493</a>) at 1/5000 dilution

Lane 1:

Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

SDHA knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

Western blot - Human SDHA knockout HEK-293 cell line (<a href='/en-us/products/cell-lines/human-sdha-knockout-hek-293-cell-line-ab261853'>ab261853</a>)

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 72 kDa

false

Next Generation Sequencing - Human SDHA knockout HEK-293 cell lysate (AB261657)
  • NGS

Lab

Next Generation Sequencing - Human SDHA knockout HEK-293 cell lysate (AB261657)

X = 1 bp insertion

Key facts

Cell type

HEK-293

Species or organism

Human

Tissue

Kidney

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 99%

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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
SDHA
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation 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.

Succinate dehydrogenase complex flavoprotein subunit A (SDHA) also known as complex II Fp or SDH2 plays an important role in the mitochondrial electron transport chain and the tricarboxylic acid (TCA) cycle. It functions as a flavoprotein oxidoreductase catalyzing the oxidation of succinate to fumarate. With a molecular mass of approximately 72 kDa SDHA is expressed in the inner mitochondrial membrane of eukaryotic cells where it is a core component of the succinate dehydrogenase complex (SDHC). The complex is essential for cellular respiration and energy production.
Biological function summary

SDHA participates in the TCA cycle by accepting electrons from succinate which it donates to the coenzyme Q in the electron transport chain. This essential role connects SDHA to the regulation of ATP production in cells. SDHA operates as part of the larger succinate dehydrogenase (SDH) complex which includes other subunits such as SDHB SDHC and SDHD. This structurally integrated multisubunit complex influences mitochondrial integrity and cellular energy homeostasis.

Pathways

SDHA is deeply involved in the TCA cycle and oxidative phosphorylation pathway. As a part of these pathways it links to other critical enzymes such as fumarase and aconitase working in concert to drive the conversion of biochemical fuel into usable cellular energy. Its interactions with coenzyme Q and cytochrome complex enzymes are important for electron flow and proton gradient formation across the mitochondrial membrane. Such interactions are central to cellular respiration and energy generation.

Mutations in SDHA correlate with various mitochondrial diseases and cancer syndromes. Specifically SDHA mutations have an association with Leigh syndrome and certain types of mitochondrial complex II deficiency. These mutations disrupt the function of the SDH complex causing metabolic imbalances and energy production issues. Furthermore the integral interaction of SDHA with other SDH subunits means that alterations can impact this entire enzymatic complex with implications for cellular respiration and disease progression.
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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

Assay Kits

AB109908

Complex II Activity Assay Kit (Colorimetric)

Functional Studies - Complex II Activity Assay Kit (Colorimetric) (AB109908)

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Primary Antibodies

AB14715

Anti-SDHA antibody [2E3GC12FB2AE2]

KO Validated

Western blot - Anti-SDHA antibody [2E3GC12FB2AE2] (AB14715)

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Proteins & Peptides

AB226268

Recombinant Human SDHA protein (Tagged)

Mass Spectrometry - Recombinant Human SDHA protein (Tagged) (AB226268)

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