|Product Name||Anti-CEACAM5/Cd66e Monoclonal 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||Mouse IgG monoclonal antibody for CEACAM5/Cd66e detection. Tested with WB, IHC-P in Human. No cross reactivity with other proteins.|
|Cite This Product||Anti-CEACAM5/Cd66e Monoclonal Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # MA1023)|
|Immunogen||Carcinoembryonic antigen-related cell adhesion molecule 5 (CEA) isolated from a human colon adenocarcinoma cell line.|
Assay Dilutions Overview
Western blot, 0.5-1μg/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-Mouse IgG (EK1001) for Western blot, and HRP Conjugated anti-Mouse IgG Super Vision Assay Kit (SV0001-1) for IHC(P).
Images And Assay Conditions
Figure 1. Western blot analysis of CEACAM5/Cd66e using anti- CEACAM5/Cd66e antibody (MA1023).
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: Recombinant Human CEACAM5/Cd66e Protein 10ng
Lane 2; Recombinant Human CEACAM5/Cd66e Protein 5ng
Lane 3: Recombinant Human CEACAM5/Cd66e Protein 2.5ng
Lane 4: Recombinant Human CEACAM5/Cd66e Protein 1.25ng
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 mouse anti- CEACAM5/Cd66e antigen affinity purified monoclonal antibody (Catalog # MA1023) 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-mouse 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 # EK1001) with Tanon 5200 system.
Figure 2. IHC analysis of CEACAM5/Cd66e using anti-CEACAM5/Cd66e antibody (MA1023).
CEACAM5/Cd66e was detected in paraffin-embedded section of human intestinal cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-CEACAM5/Cd66e Antibody (MA1023) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Carcinoembryonic antigen-related cell adhesion molecule 5|
|Tissue Specificity||Expressed in columnar epithelial and goblet cells of the colon (at protein level) (PubMed:10436421). 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|
|Subcellular Localization||Cell membrane|
*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, intracellular signaling and tumor progression (PubMed:2803308, PubMed:10910050, PubMed:10864933). Mediates homophilic and heterophilic cell adhesion with other carcinoembryonic antigen-related cell adhesion molecules, such as CEACAM6 (PubMed:2803308). Plays a role as an oncogene by promoting tumor progression; induces resistance to anoikis of colorectal carcinoma cells|
|Background||Carcinoembryonic antigen is a complex immunoreactive glycoprotein with a molecular weight of 180,000 comprising 60% carbohydrate. It is found in adenocarcinomas of endodermally derived digestive system epithelia and in fetal colon.Carcinoembryonic antigen is one of the most widely used tumor markers in serum immunoassay determinations of carcinoma.|
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