Src Overexpression Lysate

Name: Src Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-04722

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-04722

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Product Datasheet / COA / SDS

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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 Src Overexpression Lysate

Src Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC211858

Accession#: NM_198291

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

59.7 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 Src Overexpression Lysate

Western Blot: Src Overexpression Lysate [NBP2-04722]

Western Blot: Src Overexpression Lysate (Adult Normal) [NBP2-04722] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Src.

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: Src

Src is a protein tyrosine kinase known to regulate cellular adhesion. Composed of three major domains; SH2, SH3 and a kinase catalytic domain, Src can be switched from an inactive to an active state though control of its phosphorylation state or through protein-protein interaction. Phosphorylation of Y529 by Csk and Chk inactivates Src, while dephosphorylation of Y529 will modify its conformation to an open active state (1-3). Mutation of Y529 leads to Scr overactivity. Several cancers including colon and breast cancer have been associated with an increase of Src activity (4).

Long Name

v-src Sarcoma [Schmidt-Ruppin A-2] Viral Oncogene Homolog

Alternate Names

ASV, c-Src, RSVgp4

Gene Symbol

SRC

Additional Src Products

Product Documents for Src Overexpression Lysate

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Src 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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