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AB261853

Human SDHA knockout HEK-293 cell line

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SDHA KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 99%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
4 Images
Western blot - Human SDHA knockout HEK-293 cell line (AB261853)
  • WB

Lab

Western blot - Human SDHA knockout HEK-293 cell line (AB261853)

ab198493 was shown to specifically react with SDHA in wild-type HEK-293 cells as signal was lost in SDHA knockout cells. 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° 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 (ab261853)

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 line (AB261853)
  • WB

Supplier Data

Western blot - Human SDHA knockout HEK-293 cell line (AB261853)

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° 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 (ab261853)

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

Western blot - Human SDHA knockout HEK-293 cell line (AB261853)
  • WB

Supplier Data

Western blot - Human SDHA knockout HEK-293 cell line (AB261853)

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° 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 (ab261853)

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

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Next Generation Sequencing - Human SDHA knockout HEK-293 cell line (AB261853)
  • NGS

Lab

Next Generation Sequencing - Human SDHA knockout HEK-293 cell line (AB261853)

X = 1 bp insertion

Key facts

Cell type

HEK-293

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

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

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Primary Antibodies

AB14715

Anti-SDHA antibody [2E3GC12FB2AE2]

KO Validated

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

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Assay Kits

AB109908

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Primary Antibodies

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Anti-SOD2/MnSOD antibody [EPR2560Y]

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Western blot - Anti-SOD2/MnSOD antibody [EPR2560Y] (AB68155)

  • 5
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Reactivity data

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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.

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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
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.

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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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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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