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PRAS40 Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-06037

Novus Biologicals, part of Bio-Techne
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NBP2-06037

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for PRAS40 Overexpression Lysate

PRAS40 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC200919

Accession#: NM_032375

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

27.2 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 PRAS40 Overexpression Lysate

Western Blot: PRAS40 Overexpression Lysate [NBP2-06037]

Western Blot: PRAS40 Overexpression Lysate [NBP2-06037]

Western Blot: PRAS40 Overexpression Lysate (Adult Normal) [NBP2-06037] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PRAS40.

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: PRAS40

Proline Rich Akt Substrate (PRAS40) is a 40 kDa substrate of Akt. Akt and PRAS40 can be found in the phosphoinositide 3 kinase (PI3K) pathway, which plays a role in glucose uptake, cell growth, and apoptosis inhibition. PRAS40 is a 14-3-3 binding protein that reacts with insulin, but whose precise function is not yet known. It may bind SH3 and WW domain containing proteins resulting in a change of function. Activated Akt phosphorylates PRAS40 on threonine 246. Mutation of PRAS40 threonine 246 has been shown to be apoptotic.

Long Name

40 kDa Proline-rich Akt1 Substrate

Alternate Names

AKT1S1, Lobe

Gene Symbol

AKT1S1

Additional PRAS40 Products

Product Documents for PRAS40 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 PRAS40 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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