MIC0102

antibody from Invitrogen Antibodies

Targeting: CEACAM5 CD66e, CEA

Antibody data

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Validation data

Product number: MIC0102 - Provider product page
Provider: Invitrogen Antibodies
Product name: CEA Monoclonal Antibody (1106)
Antibody type: Monoclonal
Antigen: Purifed from natural sources
Description: The predicted MW of CEA is ~77kD, but by Western blot MIC0102 detects CEA with varying degrees of glycosylation at ~77-180kD.
Antibody clone number: 1106
Concentration: 5 mg/mL

CEACAM5 protein structure - MIC0102 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Carcinoembryonic Antigen (CEA) was performed by loading 20 µg of BxPC-3, HeLa, and HepG2 whole cell lysates per well and 5 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% milk in TBST for 1 hour at room temperature. CEA (with varying degrees of glycosylation) was detected at ~77-180 kD using a CEA monoclonal antibody (Product # MIC0102) at a dilution of 1 µg/mL in 5% milk in TBST overnight at 4C on a rocking platform, followed by a goat anti-mouse IgG HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico substrate (Product # 34080).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-CEA Monoclonal Antibody (1106) (Product # MIC0102) and a 160 kDa band corresponding to Carcinoembryonic antigen-related cell adhesion molecule 5 was observed across cell lines tested. Membrane enriched extracts (30 µg lysate) of MCF7 (Lane 1), HCT 116 (Lane 2), SW480 (Lane 3), HT-29 (Lane 4), HT-29 treated with IL-6,100 ng/mL, 48 Hrs. (Lane 5), MDA-MB-231 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 Dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Carcinoembryonic Antigen (CEA, green) in BxPC-3 cells. Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, and blocked with 0.3% BSA in PBS, each for 15 minutes at room temperature. Cells were stained with a CEA monoclonal antibody (Product # MIC0102) at a dilution of 10 µg/mL in blocking buffer for 1 hour at room temperature, and then incubated with a goat anti-mouse IgG Superclonal secondary antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:1000 for 1 hour at room temperature. Nuclei (blue) were stained with Hoechst nuclear stain. Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Carcinoembryonic antigen-related cell adhesion molecule 5 was performed using 70% confluent log phase HT-29 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with CEA Monoclonal Antibody (1106) (Product # MIC0102) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cell membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60x magnification.