This Human EDEM2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of EDEM2 protein (Cat: 15687-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human EDEM2 (AAH01371.1) (Met1-Lys492) was expressed with a polyhistidine tag at the C-terminus.
The recombinant human EDEM2 consists of 482 amino acids and predicts a molecular mass of 54 KDa. It migrates as an approximately 53-58 KDa band in SDS-PAGE under reducing conditions.
Human EDEM2 HEK293 Overexpression Lysate: 使用指南
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.
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.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human EDEM2 HEK293 Overexpression Lysate: 别称
Human bA4204.1 Overexpression Lysate; Human C20orf31 Overexpression Lysate; Human C20orf49 Overexpression Lysate; Human UNQ573/PRO1135 Overexpression Lysate
EDEM2, also known as C2orf31, belongs to a family of proteins involved in ER-associated degradation (ERAD) of glycoproteins. In the endoplasmic reticulum (ER), misfolded proteins are retrotranslocated to the cytosol and degraded by the proteasome. Early in this pathway, a proposed lumenal ER lectin, EDEM, recognizes misfolded glycoproteins in the ER, disengages the nascent molecules from the folding pathway, and facilitates their targeting for disposal. In humans there are a total of three EDEM homologs. The amino acid sequences of these proteins are different from other lectins but are closely related to the Class I mannosidases (family 47 glycosidases). EDEM2 is one of the EDEM homologs. Overexpression of EDEM2 accelerates the degradation of misfolded alpha1-antitrypsin, indicating that the protein is involved in ERAD.
ER degradation enhancer, mannosidase alpha-like 2
Mast SW. et al., 2005, Glycobiology. 15 (4): 421-36.
Olivari S. et al., 2005, J Biol Chem. 280 (4): 2424-8.