Human CEACAM3 HEK293 Overexpression Lysate

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Human CEACAM3 HEK293 Overexpression Lysate: Product Information

Product Description
This Human CEACAM3 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CEACAM3 protein (Cat: 11933-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human CEACAM3 (NP_001806.2) extracellular domain (Met 1-Gly 155) was expressed, with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human CEACAM3 consists of 132 amino acids and predictes a molecular mass of 14.5 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhCEACAM3 is approximately 18-23 kDa due to glycosylation.

Human CEACAM3 HEK293 Overexpression Lysate: Usage Guide

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 CEACAM3 HEK293 Overexpression Lysate: Alternative Names

Human CD66D Overexpression Lysate; Human CEA Overexpression Lysate; Human CGM1 Overexpression Lysate; Human MGC119875 Overexpression Lysate; Human W264 Overexpression Lysate; Human W282 Overexpression Lysate

CEACAM3 Background Information

CeACAM3 (CD66d), a member of carcinoembryonic antigen family, is a granulocyte-specific receptor involved in the opsonin-independent recognition of several bacterial pathogens. There are four members in this family: CD66a, CD66b, CD66c, and CD66d. Members of CEACAM family are widely expressed especially on human neutrophils, and, depending on the tissue, capable of regulating diverse functions including tumor promotion, tumor suppression, angiogenesis, and neutrophil activation. Abnormal overexpression and downregulation of some CEACAMs have been described in tumor cells. Monoclonal antibodies grouped in the CD66 cluster recognize CEACAM members. Ectopic CD66 expression is commonly detected in B-cell lineage acute lymphoblastic leukemia (ALL). CEACAM3 mediates phagocytosis depends on the integrity of an ITAM-like sequence within the cytoplasmic domain of CEACAM3. CEACAM3 is characterized by rapid stimulation of the GTPase Rac.
Full Name
carcinoembryonic antigen related cell adhesion molecule 3
References
  • Schmitter T, et al. (2007) The Granulocyte Receptor Carcinoembryonic Antigen-Related Cell Adhesion Molecule 3 (CEACAM3) Directly Associates with Vav to Promote Phagocytosis of Human Pathogens. The journal of immunology. 178: 3797-805.
  • Swanson KV, et al. (2001) CEACAM is not necessary for Neisseria gonorrhoeae to adhere to and invade female genital epithelial cells. Cellular Microbiology. 3 (10): 681-91.
  • Hasselbalch HC, et al. (2011) High expression of carcinoembryonic antigen-related cell adhesion molecule (CEACAM) 6 and 8 in primary myelofibrosis. Leukemia Research.
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