Anti-CEACAM1 antibody [EPR4049]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM1 antibody [EPR4049] (AB108397)

Unpurified ab108397 at 1/50 dilution staining CEACAM1 in Human kidney tissue by Immunohistochemistry, Paraffin-embedded tissue.

Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM1 antibody [EPR4049] (AB108397)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human clear cell carcinoma of kidney tissue sections labeling CEACAM1 with Purified ab108397 at 1 : 80 dilution (3.3 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IP

Unknown

Immunoprecipitation - Anti-CEACAM1 antibody [EPR4049] (AB108397)

ab108397 (purified) at 1 : 20 dilution (1.5μg) immunoprecipitating CEACAM1 in SW480 whole cell lysate.
Lane 1 (input) : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab108397 & SW480 whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab108397 in SW480 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-CEACAM1 antibody [EPR4049] (ab108397)

Predicted band size: 57 kDa

false

• WB

Lab

Western blot - Anti-CEACAM1 antibody [EPR4049] (AB108397)

Lanes 1-4 : Merged signal (red and green). Green - ab108397 observed at 100-180 kDa. Red - loading control ab8245 observed at 36 kDa.

ab108397 Anti-CEACAM1 antibody [EPR4049] was shown to specifically react with CEACAM1 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267047 (knockout cell lysate ab257389) was used. Wild-type and CEACAM1 knockout samples were subjected to SDS-PAGE. ab108397 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CEACAM1 antibody [EPR4049] (ab108397) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

CEACAM1 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human CEACAM1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-ceacam1-knockout-a549-cell-line-ab267047'>ab267047</a>)

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 57 kDa

Observed band size: 100-180 kDa

false

• WB

Lab

Western blot - Anti-CEACAM1 antibody [EPR4049] (AB108397)

Lanes 1-4 : Merged signal (red and green). Green - ab108397 observed at 100-180 kDa. Red - loading control ab8245 observed at 36 kDa.

ab108397 Anti-CEACAM1 antibody [EPR4049] was shown to specifically react with CEACAM1 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267046 (knockout cell lysate ab257388) was used. Wild-type and CEACAM1 knockout samples were subjected to SDS-PAGE. ab108397 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CEACAM1 antibody [EPR4049] (ab108397) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

CEACAM1 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human CEACAM1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-ceacam1-knockout-a549-cell-line-ab267046'>ab267046</a>)

Lane 3:

A549 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 29 kDa,57 kDa

Observed band size: 100-180 kDa,30 kDa

false

• WB

Lab

Western blot - Anti-CEACAM1 antibody [EPR4049] (AB108397)

The smear and multiple bands are consistent with what have been described in the literature PMID : 17071610, 26448580 and 30050594, due to a variety of splice variants and/or differential glycosylation

All lanes:

Western blot - Anti-CEACAM1 antibody [EPR4049] (ab108397) at 1/10000 dilution

Lane 1:

A549 (Human lung carcinoma epithelial cell) whole cell lysates at 20 µg

Lane 2:

SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 3:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 57 kDa

Observed band size: 100-180 kDa

false