MA5-23985

antibody from Invitrogen Antibodies

Targeting: CEACAM1 BGP, BGP1, CD66a

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Recommended by provider)

Antibody Data

Product number

MA5-23985

Provider

Invitrogen Antibodies

Product name

CEACAM1 Monoclonal Antibody (283340)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

In direct ELISAs and Western blots, this antibody does not cross-react with recombinant human (rh) CD31, rhICAM-1, -2, -3, recombinant mouse MAdCAM-1, or rhVCAM-1. Reconstitute at 0.5 mg/mL in sterile PBS.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

283340

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-CEACAM1 Monoclonal Antibody (Product # MA5-23985) and a 130 kDa band corresponding to CEACAM1 was observed across cell lines except PC-3 and Caki-1 which are reported to be negative. Membrane enriched extracts (30 µg lysate) of Hep G2 (Lane 1), COLO 205 (Lane 2), PC-3 (Lane 3) and Caki-1 (Lane 4) 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 ug/ml) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of CEACAM1 was performed using COLO 205 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 1 hour at room temperature. The cells were labeled with CEACAM1 Monoclonal Antibody (283340) (Product # MA5-23985) at 5 µg/mL concentration in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing plasma membrane and cytoplasm localization. Panel e represents PC-3 cells having no expression of CEACAM1. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.