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Human EGF HEK293 Overexpression Lysate

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Specification

Cat. No.

NSC896

Category

Neural Stem Cell Growth Factors

Product Type

Lysates

Product Name

Human EGF HEK293 Overexpression Lysate

Size

300 µg

Price

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Synonyms

Human Epidermal Growth Factor Overexpression Lysate; Human HOMG4 Overexpression Lysate; Human URG Overexpression Lysate

Species

Human

Molecule Mass

The recombinant human Fc/EGF chimera is a disulfide-linked homodimeric protein. The reduced monomer consists of 290 amino acids and has a calculated molecular mass of 33 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhFc/EGF monomer is approximately 37 kDa due to the glycosylation.

Stability & Storage

Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.

Product Description

This Human EGF overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of EGF protein (Cat: 10605-H01H) from the overexpression lysate was verified.

Expression Host

HEK293 Cells

Sequence Information

A DNA sequence encoding the mature soluble form of human EGF (NP_001954.2) (Asn 971-Arg 1023) was fused with the Fc region of human IgG1 at the N-terminus.

Preparation Method

Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.

Lysis Buffer

Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.

Recommend Usage

1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.

Sample Buffer

1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).

Application

Western Blot (WB)Optimal dilutions/concentrations should be determined by the end user.

Related Pathways

EGFR Signaling Pathway Jak-Stat Signaling Pathway

All of our services and products are intended for preclinical research use only and cannot be used to diagnose, treat or manage patients.