Anti-CEACAM1 + CEACAM3 + CEACAM6 antibody [YTH71.3]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM1 + CEACAM3 + CEACAM6 antibody [YTH71.3] (AB104450)

Immunohistochemical analysis of paraffin-embedded Human liver tissue using ab104450 at a 1/100 dilution followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in bile ducts of human liver.
The section was incubated with ab104450 for 30 mins at room temperature, followed by anti-rat IgG antibody (ab102248) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument
Hematoxylin was used as a counter stain.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM1 + CEACAM3 + CEACAM6 antibody [YTH71.3] (AB104450)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue using ab104450 at a 1/100 dilution followed by a ready to use secondary antibody LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in blood vessel endothelium of human kidney.
The section was incubated with ab104450 for 30 mins at room temperature, followed by anti-rat IgG antibody (ab102248) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument
Hematoxylin was used as a counter stain.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• Flow Cyt

Lab

Flow Cytometry - Anti-CEACAM1 + CEACAM3 + CEACAM6 antibody [YTH71.3] (AB104450)

Flow cytometric analysis of HeLa (human cervical adenocarcinoma epithelial cell, Left) / HepG2 (human hepatocellular carcinoma epithelial cell, Right) cells labelling CEACAM1 + CEACAM3 + CEACAM6 with ab104450 at 1/1000 dilution (0.1&micor;g) followed by a secondary antibody Goat F(ab)2 Anti-Rat IgG Fc (Alexa Fluor^® 488, ab150161)used at a 1/2000 dilution (Red) compared with an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Negative control : HeLa (PMID : 11580753)
Gated on viable cells.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-CEACAM1 + CEACAM3 + CEACAM6 antibody [YTH71.3] (AB104450)

Immunohistochemical analysis of 10% PFA-fixed, 0.1% Triton X-100 permeabilized frozen Human colon (fresh) tissue labeling CEACAM1 + CEACAM3 + CEACAM6 with ab104450 at 1/200 dilution followed by ab150167 Goat Anti-Rat IgG H&L (Alexa Fluor^® 647) preadsorbed at 1/1000 dilution.

Confocal image showing positive staining on human colon. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150167 Goat Anti-Rat IgG H&L (Alexa Fluor^® 647) preadsorbed at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CEACAM1 + CEACAM3 + CEACAM6 antibody [YTH71.3] (AB104450)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 cells labelling CEACAM1 + CEACAM3 + CEACAM6 with ab104450 at 1/250 dilution followed by secondary antibody ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (green).

ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used as a counterstain at a 1/200 dilution followed by a secondary antibody ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) used at a 1/1000 dilution (red).

Confocal image showing membranous staining in HepG2 cell line.
Negative control : HeLa (PMID : 11580753).

Nuclear DNA was labelled with DAPI (shown in blue).

• WB

Lab

Western blot - Anti-CEACAM1 + CEACAM3 + CEACAM6 antibody [YTH71.3] (AB104450)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : HeLa (PMID : 11580753)
The expression profile observed is consistent with what has been described in the literature (PMID : 32064046 and PMID : 24040308).

All lanes:

Western blot - Anti-CEACAM1 + CEACAM3 + CEACAM6 antibody [YTH71.3] (ab104450) at 1/1000 dilution

Lane 1:

Untreated HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HepG2 whole cell lysate treated with PNGase F at 20 µg

Lane 3:

Untreated A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

A549 whole cell lysate treated with PNGase F at 20 µg

Lane 5:

HT-29 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 6:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 7:

BxPC-3 (human pancreas adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/20000 dilution

Observed band size: 40-160 kDa

false

Exposure time: 5s

• WB

Lab

Western blot - Anti-CEACAM1 + CEACAM3 + CEACAM6 antibody [YTH71.3] (AB104450)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This blot was developed using a high sensitivity ECL substrate.
This antibody has low cross-reactivity with human CEACAM3.

All lanes:

Western blot - Anti-CEACAM1 + CEACAM3 + CEACAM6 antibody [YTH71.3] (ab104450) at 1/1000 dilution

All lanes:

His-tagged human CEACAM3 recombinant protein(<a href='/en-us/products/proteins-peptides/recombinant-human-ceacam3-protein-his-tag-ab276444'>ab276444</a>) at 20 ng

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/10000 dilution

Observed band size: 18 kDa

true

Exposure time: 3min

• WB

Lab

Western blot - Anti-CEACAM1 + CEACAM3 + CEACAM6 antibody [YTH71.3] (AB104450)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CEACAM1 + CEACAM3 + CEACAM6 antibody [YTH71.3] (ab104450) at 1/1000 dilution

Lane 1:

His-tagged human CEACAM1 recombinant protein at 10 ng

Lane 2:

His-tagged human CEACAM5 recombinant protein at 10 ng

Lane 3:

His-tagged human CEACAM6 recombinant protein at 10 ng

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/10000 dilution

Observed band size: 40-160 kDa

false

Exposure time: 92s