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Pyruvate Dehydrogenase E1-alpha subunit Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-14241

Novus Biologicals, part of Bio-Techne
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NBL1-14241

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Pyruvate Dehydrogenase E1-alpha subunit Overexpression Lysate

Pyruvate Dehydrogenase E1-alpha subunit Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC201831

Accession#: NM_000284

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

TMW

40.2 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for Pyruvate Dehydrogenase E1-alpha subunit Overexpression Lysate

Western Blot: Pyruvate Dehydrogenase E1-alpha subunit Overexpression Lysate [NBL1-14241]

Western Blot: Pyruvate Dehydrogenase E1-alpha subunit Overexpression Lysate [NBL1-14241]

Western Blot: Pyruvate Dehydrogenase E1-alpha subunit Overexpression Lysate (Adult Normal) [NBL1-14241] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Pyruvate Dehydrogenase E1-alpha subunit.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: Pyruvate Dehydrogenase E1-alpha subunit

The pyruvate dehydrogenase complex is a nuclear-encoded mitochondrial matrix multienzyme complex that provides the primary link between glycolysis and the tricarboxylic acid (TCA) cycle by catalyzing the irreversible conversion of pyruvate into acetyl-CoA. The PDH complex is composed of multiple copies of 3 enzymes: E1 (PDHA1); dihydrolipoyl transacetylase (DLAT; MIM 608770) (E2; EC 2.3.1.12); and dihydrolipoyl dehydrogenase (DLD; MIM 238331) (E3; EC 1.8.1.4). The E1 enzyme is a heterotetramer of 2 alpha and 2 beta subunits. The E1-alpha subunit contains the E1 active site and plays a key role in the function of the PDH complex (Brown et al., 1994 [PubMed 7853374]).[supplied by OMIM]. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

Alternate Names

EC 1.2.4.1, PDHA, PDHE1-A type I, PHE1APDHCE1A, pyruvate dehydrogenase (lipoamide) alpha 1, pyruvate dehydrogenase complex, E1-alpha polypeptide 1, pyruvate dehydrogenase E1 component subunit alpha, somatic form, mitochondrial

Gene Symbol

PDHA1

Additional Pyruvate Dehydrogenase E1-alpha subunit Products

Product Documents for Pyruvate Dehydrogenase E1-alpha subunit Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for Pyruvate Dehydrogenase E1-alpha subunit Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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