|Applications||ELISA, IHC, WB|
|Product Name||Anti-CEACAM5 Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Carcinoembryonic antigen-related cell adhesion molecule 5(CEACAM5) detection. Tested with WB, IHC-P, ELISA in Human.|
|Cite This Product||Anti-CEACAM5 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # RP1018)|
|Immunogen||Insect cells-derived human CEA recombinant protein(Position: full).|
Assay Dilutions Overview
Immunohistochemistry(Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Western blot, 0.1-0.5μg/ml, Human
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P).
Images And Assay Conditions
Figure 1. Western blot analysis of CEA using anti- CEA antibody (RP1018).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human SW620 whole cell lysates,
Lane 2: human Caco-2 whole cell lysates,
Lane 3: mouse small intestine tissue lysates,
Lane 4: mouse stomach tissue lysates,
Lane 5: mouse lung tissue lysates,
Lane 6: mouse liver tissue lysates,
Lane 7: mouse NIH3T3 whole cell lysates,
Lane 8: mouse HEPA1-6 whole cell lysates,
Lane 9: mouse SP20 whole cell lysates,
Lane 10: rat RH35 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- CEA antigen affinity purified polyclonal antibody (Catalog # RP1018) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CEA at approximately 120-200KD. The expected band size for CEA is at 77KD.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Carcinoembryonic antigen-related cell adhesion molecule 5|
|Tissue Specificity||Found in adenocarcinomas of endodermally derived digestive system epithelium and fetal colon.|
|Alternative Names||Carcinoembryonic antigen-related cell adhesion molecule 5;Carcinoembryonic antigen;CEA;Meconium antigen 100;CD66e;CEACAM5;CEA;|
|Subcellular Localization||Cell membrane ; Lipid-anchor, GPI-anchor .|
|Molecular Weight||76795 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Cell surface glycoprotein that plays a role in cell adhesion and in intracellular signaling. Receptor for E.coli Dr adhesins. .|
|Background||CEA(Carcinoembryonic antigen) is a glycoprotein involved in cell adhesion. It is normally produced during fetal development, but the production of CEA stops before birth. Therefore, it is not usually present in the blood of healthy adults, although levels are raised in heavy smokers. CEA is a glycosyl phosphatidyl inositol(GPI)-cell surface anchored glycoprotein whose specialized sialofucosylated glycoforms serve as functional colon carcinoma L-selectin and E-selectin ligands, which may be critical to the metastatic dissemination of colon carcinoma cells. CEA and related genes make up the CEA family belonging to the immunoglobulin superfamily. In humans, the carcinoembryonic antigen family consists of 29 genes, 18 of which are normally expressed.|
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1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,