Category
Neural Stem Cell Growth Factors
Product Name
Human L1CAM HEK293 Overexpression Lysate
Synonyms
Human CAML1 Overexpression Lysate; Human CD171 Overexpression Lysate; Human HSAS Overexpression Lysate; Human HSAS1 Overexpression Lysate; Human MASA Overexpression Lysate; Human MIC5 Overexpression Lysate; Human N-CAM-L1 Overexpression Lysate; Human N-CAML1 Overexpression Lysate; Human NCAM-L1 Overexpression Lysate; Human S10 Overexpression Lysate; Human SPG1 Overexpression Lysate
Molecule Mass
The recombinant human CD171 consists of 1112 amino acids after removal of the signal peptide and predicts a molecular mass of 125 kDa. As a result of glycosylation, the apparent molecular mass of rhCD171 is approximately 160-200 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 L1CAM overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of L1CAM protein (Cat: 10140-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the extracellular domain of human CD171 (NP_000416.1) (Met 1-Glu 1120) was fused with a polyhistidine tag 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.