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Human CD166/ALCAM HEK293 Overexpression Lysate

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Specification

Cat. No.

NSC567

Category

Neural Stem Cell Growth Factors

Product Type

Lysates

Product Name

Human CD166/ALCAM HEK293 Overexpression Lysate

Size

300 µg

Price

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Synonyms

Human CD166 Overexpression Lysate; Human MEMD Overexpression Lysate

Species

Human

Molecule Mass

The recombinant human ALCAM consists of 510 amino acids and has a calculated molecular mass of 57.4 kDa. As a result of glycosylation, human ALCAM migrates as an approximately 78.3 kDa band in SDS-PAGE under reducing conditions.

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 CD166/ALCAM overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD166/ALCAM protein (Cat: 10045-H08H) from the overexpression lysate was verified.

Expression Host

HEK293 Cells

Sequence Information

A DNA sequence encoding the extracellular domain (Met1-Ala526) of human ALCAM (NP_001618.2) was expressed with the 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.