Human N-Cadherin HEK293 Overexpression Lysate

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Human N-Cadherin HEK293 Overexpression Lysate

Specification
Cat. No.
NSC1350
Category
Neural Stem Cell Markers
Product Type
Lysates
Product Name
Human N-Cadherin HEK293 Overexpression Lysate
Size
300 µg
Price
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Synonyms
Human CD325 Overexpression Lysate; Human CDHN Overexpression Lysate; Human CDw325 Overexpression Lysate; Human NCAD Overexpression Lysate
Species
Human
Molecule Mass
The secreted recombinant human CDH2 is a disulfide-linked homodimeric protein. The reduced monomer comprises 813 amino acids and has a predicted molecular mass of 89.9 kDa. As a result of glycosylation, it migrates as an approximately 114 and 119 kDa band 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 Human N-Cadherin overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of N-Cadherin protein (Cat: 11039-H03H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human CDH2 (NP_001783.2) (Met 1-Ala 724) was fused with the C-terminal polyhistidine-tagged 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.
Related Pathways
Canonical Wnt Pathway
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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