711088

antibody from Invitrogen Antibodies

Targeting: CEACAM1 BGP, BGP1, CD66a

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

711088

Provider

Invitrogen Antibodies

Product name

CEACAM1 Recombinant Polyclonal Antibody (10HCLC)

Antibody type

Polyclonal

Antigen

Other

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

10HCLC

Vial size

100 µg

Concentration

0.5 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 analysis was performed on Membrane extracts (30 µg lysate) of Hep G2 (Lane 1) and HT-29 (Lane 2). The blots were probed with Anti-Ceacam1 Recombinant Rabbit Polyclonal Antibody (Product # 711088, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Three isoforms at 150, 35 and 27 kDa band corresponding to Ceacam1 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-CEACAM1 Recombinant Polyclonal Antibody (10HCLC) (Product # 711088) and a 140 kDa band corresponding to CEACAM1 was observed in HepG2 and no band was seen in PC-3 which is a low expression model. Whole cell extracts (50 µg lysate) of Hep G2 (Lane 1), PC-3 (Lane 2) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 (1:500) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:10000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-CEACAM1 Antibody (10HCLC), ABfinity Rabbit Oligoclonal (Product # 711088) and a 150kDa band corresponding to CEACAM1 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), COLO 205 (Lane 2), PC-3 (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).Expression of CEACAM1 was found to be higher in Hep G2 cells as compared to COLO-205 and PC-3 cells.