Rat CD166/ALCAM HEK293 Overexpression Lysate

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Rat CD166/ALCAM HEK293 Overexpression Lysate

Specification
Cat. No.
NSC571
Category
Neural Stem Cell Growth Factors
Product Type
Lysates
Product Name
Rat CD166/ALCAM HEK293 Overexpression Lysate
Size
300 µg
Price
Inquiry
Species
Rat
Molecule Mass
The recombinant rat ALCAM/Fc is a disulfide-linked homodimer. The reduced monomer comprises 741 amino acids and has a predicted molecular mass of 83.1 kDa. The apparent molecular mass of the protein is approximately 94 kDa in SDS-PAGE under reducing conditions due to 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 Rat 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: 80221-R02H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the rat ALCAM (O35112) (Met1-Lys527) was expressed with the Fc region of human IgG1 at the C-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.
All of our services and products are intended for preclinical research use only and cannot be used to diagnose, treat or manage patients.

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