Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

ab133633 showing negative staining in Normal tonsil tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133633).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

Immunohistochemistry analysis of paraffin-embedded (A) HEK-293T cells transfected with a CEACAM1 expression vector containing a his tag, (B) HEK-293T cells transfected with a CEACAM3 expression vector containing a his tag, (C) HEK-293T cells transfected with a CEACAM5 expression vector containing a his tag, (D) HEK-293T cells transfected with a CEACAM6 expression vector containing a his tag, (E) HEK-293T cells transfected with a CEACAM8 expression vector containing a his tag and (F) HEK-293T cells transfected with empty vector containing a his tag, sections labelling Carcino Embryonic Antigen CEA with ab133633 at 1/2000 dilution. LeicaDS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody. Sections were counterstained with Hematoxylin. The section was incubated with ab133633 for 30 mins at room temperature. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Positive staining on HEK-293T cells transfected with a CEACAM5 expression vector containing a his tag (Image C), and HEK-293T cells transfected with a CEACAM6 expression vector containing a his tag (Image D). No staining on other cell pellet tested. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. IHC Note : ab133633 is also cross-reactIve with human CEACAM6 in IHC. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133633).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

Immunohistochemical analysis of paraffin embedded Human colon carcinoma tissue labelling Carcino Embryonic Antigen CEA with ab133633 at 1/250.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133633).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human gastric carcinoma tissue sections labeling Carcino Embryonic Antigen CEA with Purified ab133633 at 1 : 3000 dilution (0.6 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, PH9. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133633).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

ab133633 showing positive staining in Stomach adenocarcinoma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133633).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

Confocal image showing membranous staining in MCF7 cell line.

Immunocytochemistry/Immunofluorescence analysis of MCF7 (human breast adenocarcinoma cell line) cells labelling carcino embryonic antigen (CEA) with ab133633 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/10000 dilution was used as the secondary antibody. The cells were counterstained with ab195889, a anti-alpha tubulin antibody at 1/200 dilution. DAPI was used as a nuclear counterstain.

Secondary antibody only control : Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133633).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling Carcino Embryonic Antigen CEA with Purified ab133633 at 1 : 3000 dilution (0.6 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using EDTA Buffer, PH9. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133633).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

ab133633 showing negative staining in Hepatocellular carcinoma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133633).

• WB

Lab

Western blot - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

This data was developed using ab133633, the same antibody clone in a different buffer formulation.

Exposure time : Lane 1 : 10 seconds; Lane 2 : 180 seconds

We recommend not to boil the sample after lysis to get desirable WB result.

All lanes:

Western blot - Anti-CEACAM5 antibody [EPCEAR7] (<a href='/en-us/products/primary-antibodies/carcino-embryonic-antigen-cea-antibody-epcear7-ab133633'>ab133633</a>)

Lane 1:

Human colon tissue lysate unboiled at 20 µg

Lane 2:

Human colon tissue lysate boiled at 20 µg

Observed band size: 180-200 kDa

false

• WB

Lab

Western blot - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133633). Western blot : Anti-CEACAM5 antibody [EPCEAR7] (ab133633) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab133633 was shown to bind specifically to CEACAM5. A band was observed at 160-180 kDa in wild-type A549 cell lysates with no signal observed at this size in CEACAM5 knockout cell line. To generate this image, wild-type and CEACAM5 knockout A549 unboiled cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-CEACAM5 antibody [EPCEAR7] (<a href='/en-us/products/primary-antibodies/carcino-embryonic-antigen-cea-antibody-epcear7-ab133633'>ab133633</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

CEACAM5 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human CEACAM5 knockout A549 cell line (ab287319)

Lane 3:

MCF7 (unboiled) cell lysate at 20 µg

Lane 4:

PANC-1 (unboiled) cell lysate at 20 µg

Observed band size: 160-180 kDa

true

• WB

Supplier Data

Western blot - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

Blocking buffer and concentration :  5% NFDM/TBST.  Diluting buffer and concentration :  5% NFDM/TBST. ab181602 was used as a loading control. We recommend not to boil the sample after lysis to get desirable WB result. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133633).

All lanes:

Western blot - Anti-CEACAM5 antibody [EPCEAR7] (<a href='/en-us/products/primary-antibodies/carcino-embryonic-antigen-cea-antibody-epcear7-ab133633'>ab133633</a>) at 1/1000 dilution

Lane 1:

BxPC-3 (Human pancreas adenocarcinoma epithelial cell) whole cell lysate boiled at 20 µg

Lane 2:

BxPC-3 (Human pancreas adenocarcinoma epithelial cell) whole cell lysate unboiled at 20 µg

Lane 3:

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

Lane 4:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate unboiled 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

Predicted band size: 76 kDa

Observed band size: 180-200 kDa

false

Exposure time: 3s

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

ab133633 showing negative staining in Skeletal muscle tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133633).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

ab133633 showing negative staining in Thyroid gland carcinoma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133633).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

ab133633 showing negative staining in Glioma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133633).

• WB

Lab

Western blot - Anti-CEACAM5 antibody [EPCEAR7] - BSA and Azide free (AB226144)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133633).

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

This antibody does not cross-react with human CEACAM1, CEACAM3, CEACAM6, and CEACAM8.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-CEACAM5 antibody [EPCEAR7] (<a href='/en-us/products/primary-antibodies/carcino-embryonic-antigen-cea-antibody-epcear7-ab133633'>ab133633</a>) at 1/1000 dilution

Lane 1:

His-tagged human CEACAM1 recombinant protein (aa 35-428) at 0.02 µg

Lane 2:

His-tagged human CEACAM3 recombinant protein (aa 35-252) at 0.02 µg

Lane 3:

His-tagged human CEACAM5 recombinant protein (aa 35-586) at 0.02 µg

Lane 4:

His-tagged human CEACAM6 recombinant protein (aa 35-320) at 0.02 µg

Lane 5:

His-tagged human CEACAM8 recombinant protein (aa 35-320) at 0.02 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 1/100000 dilution

Predicted band size: 76 kDa

Observed band size: 180 kDa

true

Exposure time: 3min