SDHD KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 1.
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Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 1
Alternative names
CBT1, CII-4, CWS3, CybS, DHSD_HUMAN, OTTHUMP00000234720, OTTHUMP00000234721, OTTHUMP00000234722, OTTHUMP00000234723, OTTHUMP00000234724, OTTHUMP00000234725, OTTHUMP00000234726, PGL, PGL1, QPs3, SDH4, Succinate dehydrogenase [ubiquinone] cytochrome b small subunit, Succinate dehydrogenase [ubiquinone] cytochrome b small subunit, mitochondrial, Succinate dehydrogenase complex subunit D, Succinate dehydrogenase complex, subunit D, integral membrane protein, Succinate dehydrogenase ubiquinone cytochrome B small subunit, Succinate-ubiquinone oxidoreductase cytochrome b small subunit, Succinate-ubiquinone reductase membrane anchor subunit, mitochondrial
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SDHD KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 1.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 1
- Concentration
Properties
- Gene name
- SDHD
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
- Zygosity
- Homozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The SDHD protein also known as succinate dehydrogenase complex subunit D serves a function in cellular respiration. This protein is a part of the succinate dehydrogenase (SDH) complex found in the inner mitochondrial membrane. This complex is known as complex II in the electron transport chain. The subunit D with a mass of about 15 kDa anchors the larger SDH complex to the membrane and is essential for its structural stability. SDHD is widely expressed in tissues with high energy demands such as the heart liver and muscles.
- Biological function summary
The SDH complex has an essential role in both the Krebs cycle and the mitochondrial electron transport chain. SDHD as part of this complex assists in the oxidation of succinate to fumarate an important step in the Krebs cycle. The electrons generated from succinate oxidation are transferred through the SDH complex to ubiquinone contributing to ATP production. The complex facilitates the coupling of the Krebs cycle to the electron transport chain highlighting SDHD’s importance in efficient energy metabolism.
- Pathways
SDHD integrates into the Krebs cycle and the electron transport chain linking these vital energy-yielding reactions. In the Krebs cycle it assists in converting succinate to fumarate contributing important intermediates and electron donors for oxidative phosphorylation. It interacts with other components of complex II such as SDHA SDHB and SDHC to facilitate its enzymatic functions. Furthermore its role in electron transport involves ubiquinone which carries electrons to complex III continuing the chain of reactions needed for energy production.
- Associated diseases and disorders
SDHD mutations have been linked with paragangliomas and pheochromocytomas both of which are neuroendocrine tumors. These conditions are associated with disrupted cellular energy metabolism caused by impaired function of the SDH complex. In these tumors mutations in SDHD can lead to a pseudohypoxic state promoting cell proliferation. The protein's dysfunction connects pathophysiologically to related SDHB SDHC and SDHA subunits with mutations in these subunits also contributing to the manifestation of such tumors.
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Sanger Sequencing - Human SDHD knockout HEK-293T cell line (ab266491)
Homozygous: 2 bp insertion in exon 1
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