Human CEACAM5 HEK293 Overexpression Lysate: Product Information
This Human CEACAM5 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CEACAM5 protein (Cat: 11077-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the mature form of human CEACAM5 (NP_004354.2) (Met 1-Ala 685) was fused with a polyhistidine tag at the C-terminus.
The secreted recombinant human CEACAM5 consists of 662 amino acids and has a predicted molecular mass of 72.8 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhCEACAM5 is approximately 100-110 kDa due to glycosylation.
Human CEACAM5 HEK293 Overexpression Lysate: Usage Guide
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).
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human CEACAM5 HEK293 Overexpression Lysate: Alternative Names
Human CD66e Overexpression Lysate; Human CEA Overexpression Lysate
CEACAM5 Background Information
CEACAM5, also known as CEA or D66e, belongs to the large CEACAM subfamily of immunoglobulin superfamily. CEACAM5 is expressed primarily by epithelial cells, and is synthesized as a glycoprotein with a MW of 18 kDa comprising 6% carbohydrate. CEACAM5 contains one Ig-like V-type domain at the N-terminus, followed by six Ig-like C2-type domain and a GPI anchor, and exists as a homodimer. CEACAM5 and CEACAM6 are overexpressed in many cancers and are associated with adhesion and invasion. CEACAM5 can mediate cell-cell adhesion through homotypic and heterotypic interactions. It functions as a homotypic intercellular adhesion molecule and serves as a widely used tumor marker, since it is expressed at higher levels in tumorous tissues than in corresponding normal tissues. CEACAM5 has also been shown to contribute to tumorigenicity by inhibiting cellular differentiation. In addition, CEACAM5 is identified as the host receptor for the Dr family of adhesins of E.Coli, and the binding of E.coli Dr adhesins leads to dissociation of the CEACAM5 homodimer.
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