PEA-15 Overexpression Lysate

Name: PEA-15 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-14273

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-14273

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for PEA-15 Overexpression Lysate

PEA-15 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC205507

Accession#: NM_003768

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

14.9 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 PEA-15 Overexpression Lysate

Western Blot: PEA-15 Overexpression Lysate [NBL1-14273]

Western Blot: PEA15 Overexpression Lysate (Adult Normal) [NBL1-14273] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PEA15.

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: PEA-15

PEA-15 (phosphoprotein enriched in astrocytes) exists in an non-phosphorylated form (N), and two phosphorylated forms, Pa and Pb. PEA-15 is an endogenous substrate for PKC, which mediates the transition from Pa to Pb. The level of PEA-15 phosphorylation changes upon depolymerization or stabilization of tubulins, indicating that PEA-15 co-localizes with microtubules. The first 80 amino acids of PEA-15 correspond to the death effector domain (DED), which is a domain found in proteins that regulate apoptotic signaling pathways. The DED domain is necessary for PEA-15 to block Ras suppression. Although PEA-15 is predominantly expressed in the central nervous system, low levels of PEA-15 are expressed in liver and kidney, and higher levels in muscle. PEA-15 is also referred to as PED, phosphoprotein enriched in diabetes, for its elevated expression in type 2 diabetic patients.

Long Name

Phosphoprotein Enriched in Astrocytes 15

Alternate Names

HMAT1, HUMMAT1H, MAT1H, PEA15, PED

Gene Symbol

PEA15

Additional PEA-15 Products

Product Documents for PEA-15 Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for PEA-15 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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