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AB300919

Human DLAT knockout A549 cell line

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DLAT KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View Alternative Names

70 kDa mitochondrial autoantigen of primary biliary cirrhosis, DLAT, DLTA, Dihydrolipoamide S Acetyltransferase, Dihydrolipoamide acetyltransferase component of pyruvate dehydrogenase complex, Dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex, E2, E2 component of pyruvate dehydrogenase complex, M2 antigen complex 70 kDa subunit, ODP2_HUMAN, PBC, PDC-E2, Pyruvate dehydrogenase complex E2 subunit, Pyruvate dehydrogenase complex component E2, dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex mitochondrial, mitochondrial

1 Images
Next Generation Sequencing - Human DLAT knockout A549 cell line (AB300919)
  • NGS

Lab

Next Generation Sequencing - Human DLAT knockout A549 cell line (AB300919)

1 bp insertion and 85 bp deletion after Pro 13 of WT protein

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Next Generation Sequencing

Disease

Carcinoma

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

What's included?

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Properties and storage information

Gene name
DLAT
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium

F-12K + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Pyruvate Dehydrogenase E2 (PDH-E2) also referred to as PDC-E2 or E2 protein is a central component of cellular metabolism. It is the inner core dihydrolipoamide acetyltransferase component of the pyruvate dehydrogenase complex. PDH-E2 facilitates the conversion of pyruvate to acetyl-CoA which is an important step in aerobic respiration. The E2 protein is expressed in the mitochondria where it actively engages in its enzymatic roles. With a molecular weight of approximately 55 kDa PDH-E2 functions through its interaction with other components of the pyruvate dehydrogenase complex.
Biological function summary

PDH-E2 plays a significant role in cellular energy production. It is an important part of the pyruvate dehydrogenase complex where its coordinated enzymatic activity drives the decarboxylation of pyruvate. This activity supplies acetyl-CoA for the citric acid cycle essential for ATP production. PDH-E2's function regulates the flow of carbon within cells linking glycolysis and the citric acid cycle making it essential for energy homeostasis. Its function integrates signals that regulate metabolic pathways helping maintain cellular energy balance.

Pathways

PDH-E2 integrates the glycolysis and citric acid cycle facilitating efficient energy production. It interacts with tightly linked enzymes in the pyruvate dehydrogenase complex important for catalyzing the conversion processes. Key related proteins include pyruvate dehydrogenase (E1) and dihydrolipoamide dehydrogenase (E3) which form the complex alongside E2. This complex is also critical in adjusting metabolic responses based on cellular energy needs efficiently managing substrate flux into the mitochondria.

Dysfunctional PDH-E2 activity associates with metabolic conditions like pyruvate dehydrogenase deficiency and primary biliary cholangitis (PBC). PDH deficiency leads to lactic acidosis and neurological dysfunction due to impaired energy production. In PBC the immune system aberrantly targets the E2 component causing liver damage and leading to cholestasis. These diseases highlight the critical role of PDH-E2 and its interaction with other mitochondrial proteins in maintaining metabolic health.

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

Product protocols

Product promise

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