MA1-17761

antibody from Invitrogen Antibodies

Targeting: CEACAM1 BGP, BGP1, CD66a

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-17761

Provider

Invitrogen Antibodies

Product name

CD66 (CEACAM) Monoclonal Antibody (D14HD11)

Antibody type

Monoclonal

Antigen

Other

Description

This antibody recognizes native human CEACAM1, 3, 4, 5, 6 protein expressed in transfected mammalian cells.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

D14HD11

Vial size

100 µg

Concentration

2 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-CEACAM Pan Monoclonal Antibody (D14HD11) (Product # MA1-17761) and a 150 kDa band corresponding to Carcinoembryonic antigen-related cell adhesion molecule 5 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of MCF7 (Lane 1), HCT 116 (Lane 2), MDA-MB-231 (Lane 3), HT-29 (Lane 4), HT-29 treated with IL-6 (100 ng/mL, 48 Hrs) (Lane 5) 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 Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry microscopy of CD66 (CEACAM) in CHO cells using CD66 (CEACAM) Monoclonal Antibody (Product # MA1-17761). CHO cells were transfected with an expression vector encoding CEACAM1 (A). Untransfected CHO parental cells served as negative control (B). Binding was visualized with a FITC-conjugated secondary antibody.

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 MCF-7 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 CEACAM Pan Monoclonal Antibody (D14HD11) (Product # MA1-17761) 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 Cytoplasm localization. Panel e represents MDA-MB-231. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of CEACAM 1,3,4,5,6 in BOSC23 cells using a CD66 (CEACAM) Monoclonal Antibody (Product # MA1-17760) at a 1.2 µg/10^6 cells dilution, detected with a PE-conjugated secondary antibody, as shown in red. Isotype control antibody is shown in black.