Anti-CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 antibody [RM1195]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 antibody [RM1195] (AB323805)

Immunohistochemical analysis of paraffin-embedded (A) 293T (human embryonic kidney epithelial cell) transfected with a CEACAM1 expression vector containing a his tag, (B) 293T transfected with a CEACAM3 expression vector containing a his tag, (C) 293T transfected with a CEACAM5 expression vector containing a his tag, (D) 293T transfected with a CEACAM6 expression vector containing a his tag, (E) 293T transfected with a CEACAM8 expression vector containing a his tag and (F) 293T cells transfected with an empty vector containing a his tag labeling CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 with ab323805 at 1/5000 (0.10 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) 293T transfected with a CEACAM1 expression vector containing a his tag, (C) 293T transfected with a CEACAM5 expression vector containing a his tag, (D) 293T transfected with a CEACAM6 expression vector containing a his tag, and (E) 293T transfected with a CEACAM8 expression vector containing a his tag. No staining on (B) 293T transfected with a CEACAM3 expression vector containing a his tag, and (F) 293T cells transfected with an empty vector containing a his tag.
The section was incubated with ab323805 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 antibody [RM1195] (AB323805)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HT-29 (human colorectal adenocarcinoma epithelial cell) cells labelling CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 with ab323805 at 1/250 (1.956 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/ml) dilution (Green).

Confocal image showing cytoplasmic and membranous staining in HT-29 cell line and no staining in HeLa cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : HeLa (PMID : 11580753).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 antibody [RM1195] (AB323805)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 with ab323805 at 1/5000 (0.10 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen.
The section was incubated with ab323805 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 antibody [RM1195] (AB323805)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 with ab323805 at 1/5000 (0.10 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon.
The section was incubated with ab323805 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 antibody [RM1195] (AB323805)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A549 (human lung carcinoma epithelial cell) cells labelling CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 with ab323805 at 1/250 (1.956 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/ml) dilution (Green).

Confocal image showing cytoplasmic and membranous staining in A549 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 antibody [RM1195] (AB323805)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 with ab323805 at 1/5000 (0.10 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : positive staining on scattered immune cells of human skeletal muscle.
The section was incubated with ab323805 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 antibody [RM1195] (AB323805)

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

Negative control : HeLa (PMID : 11580753).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-3 and 5 : 26 seconds; lane 4 : 26 seconds

All lanes:

Western blot - Anti-CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 antibody [RM1195] (ab323805) at 1/1000 dilution

Lane 1:

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

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 40-180 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 antibody [RM1195] (AB323805)

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

Low expression : Skeletal muscle

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 antibody [RM1195] (ab323805) at 1/1000 dilution

Lane 1:

Human colon tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 40-180 kDa,36 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 antibody [RM1195] (AB323805)

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

Exposure time : Lane 1 : 180 seconds; lane 2 : 103 seconds; lane 3-4 : 59 seconds

All lanes:

Western blot - Anti-CEACAM1+ CEACAM5 + CEACAM6 + CEACAM8 antibody [RM1195] (ab323805) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 20 µg

Lane 2:

Human lung cancer tissue lysate at 20 µg

Lane 3:

HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate at 20 µg

Lane 4:

SW1463 (human Large intestine; Rectum epithelial) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 40-180 kDa

false