Product Description
This Human CEACAM1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CEACAM1 protein (Cat: 10822-H03H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the extracellular domain of human CEACAM1 (NP_001020083.1) (Met 1-Gly 428) was fused with the C-terminal polyhistidine-tagged Fc region of human IgG1 at the C-terminus.
Molecule Mass
The secreted recombinant human CEACAM1/Fc chimera is a disulfide-linked homodimer. The reduced monomer comprises 642 amino acids and predicts a molecular mass of 71 kDa.
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).
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.
Human BGP Overexpression Lysate;Human BGP1 Overexpression Lysate;Human BGPI Overexpression Lysate;Human CD66a Overexpression Lysate;Human CEACAM?1 Overexpression Lysate
The carcinoembryonic-antigen-related cell-adhesion molecule (CEACAM) family of proteins has been implicated in various intercellular-adhesion and intracellular-signalling-mediated effects that govern the growth and differentiation of normal and cancerous cells. CEACAM1, also known as biliary glycoprotein I (BGP I) and CD66a, is a member of the carcinoembryonic antigen (CEA) gene family which belongs to the immunoglobulin superfamily. The highly glycosylated CEACAM1 contains one N-terminal V-type Ig-like domain and three C2-type Ig-like domains within its ECD, and one ITIM motif and a calmodulin binding site in the cytoplasmic region. CEACAM1 is a surface glycoprotein expressed on various blood cells, epithelial cells, and vascular cells. It was described as an adhesion molecule mediating cell adhesion via both homophilic and heterophilic manners, and was detected on leukocytes, epithelia, and endothelia. Studies have revealed that CEACAM1 performs actions in multiple cellular processes including tissue differentiation, angiogenesis, apoptosis, metastasis, as well as the modulation of innate and adaptive immune responses.
Immune Checkpoint Immune Checkpoint Detection: ELISA Antibodies Immune Checkpoint Detection: IHC Antibodies Immune Checkpoint Detection: ICC Antibodies Immune Checkpoint Detection: FCM Antibodies Immune Checkpoint Detection: WB Antibodies Immune Checkpoint Proteins Immune Checkpoint Targets Co-inhibitory Immune Checkpoint Targets Immunotherapy Cancer Immunotherapy Targeted Therapy References
Gray-Owen SD, et al. (2006) CEACAM1: contact-dependent control of immunity. Nat Rev Immunol. 6(6): 433-46. Gu A, et al. (2009) Role of Ceacam1 in VEGF induced vasculogenesis of murine embryonic stem cell-derived embryoid bodies in 3D culture. Exp Cell Res. 315(10): 1668-82. Wong C, et al. (2009) CEACAM1 negatively regulates platelet-collagen interactions and thrombus growth in vitro and in vivo. Blood. 113(8): 1818-28. 
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