Human SDHA knockout HEK-293 cell lysate (ab261657)
Overview
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Product name
Human SDHA knockout HEK-293 cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
HEK-293 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 99% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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Notes
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 - found here. 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.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.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.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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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab280407 - Human SDHA knockout HEK293 cell lysate 1 x 100µg ab259780 - Human wild-type HEK-293 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Gender
Female
Target
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Function
Flavoprotein (FP) subunit of succinate dehydrogenase (SDH) that is involved in complex II of the mitochondrial electron transport chain and is responsible for transferring electrons from succinate to ubiquinone (coenzyme Q). -
Pathway
Carbohydrate metabolism; tricarboxylic acid cycle; fumarate from succinate (eukaryal route): step 1/1. -
Involvement in disease
Defects in SDHA are a cause of mitochondrial complex II deficiency (MT-C2D) [MIM:252011]. A disorder of the mitochondrial respiratory chain with heterogeneous clinical manifestations. Clinical features include psychomotor regression in infants, poor growth with lack of speech development, severe spastic quadriplegia, dystonia, progressive leukoencephalopathy, muscle weakness, exercise intolerance, cardiomyopathy. Some patients manifest Leigh syndrome or Kearns-Sayre syndrome.
Defects in SDHA are a cause of Leigh syndrome (LS) [MIM:256000]. LS is a severe disorder characterized by bilaterally symmetrical necrotic lesions in subcortical brain regions.
Defects in SDHA are the cause of cardiomyopathy dilated type 1GG (CMD1GG) [MIM:613642]. CMD1GG is a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death. -
Sequence similarities
Belongs to the FAD-dependent oxidoreductase 2 family. FRD/SDH subfamily. -
Cellular localization
Mitochondrion inner membrane. - Information by UniProt
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Alternative names
- CMD1GG
- DHSA_HUMAN
- Flavoprotein subunit of complex II
see all
Associated products
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab261657 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
Use at an assay dependent concentration.
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Notes |
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WB
Use at an assay dependent concentration. |
Images
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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. -
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. -
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. -
X = 1 bp insertion
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab261657 has not yet been referenced specifically in any publications.