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Human Vimentin Baculovirus-Insect cells Overexpression Lysate

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Specification

Cat. No.

NSC332

Category

Neural Stem Cell Differentiation Markers

Product Type

Lysates

Product Name

Human Vimentin Baculovirus-Insect cells Overexpression Lysate

Size

300 µg

Price

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Synonyms

Human CTRCT30 Overexpression Lysate; Human HEL113 Overexpression Lysate

Species

Human

Molecule Mass

The secreted recombinant human VIM consists of 477 amino acids and predicts a molecular mass of 55.11 KDa.

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 Vimentin overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of Vimentin protein (Cat: 10028-H08B) from the overexpression lysate was verified.

Expression Host

Baculovirus-Insect cells

Sequence Information

A DNA sequence encoding the human VIM (Met1-Glu466) (P08670) was expressed with a C-terminal polyhistidine tag.

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.

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