Anti-EpCAM antibody [EPR20532-222] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EpCAM antibody [EPR20532-222] - BSA and Azide free (AB232539)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling EpCAM with ab213500 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab214880) Ready to use. Counter stained with Hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab214880). Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Positive staining on human breast carcinoma. The section was incubated with ab213500 at 4°C overnight. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213500).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EpCAM antibody [EPR20532-222] - BSA and Azide free (AB232539)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling EpCAM with ab213500 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membranous staining on rat colon [PMID : 15637741] is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213500).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-EpCAM antibody [EPR20532-222] - BSA and Azide free (AB232539)

This data was developed using the same antibody clone in a different buffer formulation (ab213500).

Lanes 1 - 3 : Merged signal (red and green). Green - ab213500 observed at 40 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

ab213500 was shown to react with EpCAM in A431 wild-type cells in Western blot. Loss of signal was observed when EpCAM knockout sample was used. A431 wild-type and EpCAM knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween^®) before incubation with ab213500 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-EpCAM antibody [EPR20532-222] (<a href='/en-us/products/primary-antibodies/epcam-antibody-epr20532-222-ab213500'>ab213500</a>) at 1/1000 dilution

Lane 1:

Wild-type A431 whole cell lysate at 20 µg

Lane 2:

EPCAM knockout A431 whole cell lysate at 20 µg

Lane 2:

Western blot - Human EPCAM knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-epcam-knockout-a-431-cell-line-ab261902'>ab261902</a>)

Lane 3:

HeLa whole cell lysate at 20 µg

Predicted band size: 35 kDa

Observed band size: 35 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-EpCAM antibody [EPR20532-222] - BSA and Azide free (AB232539)

Immunofluorescent analysis of 100% methanol-fixed 4T1 (Mouse mammary gland carcinoma cell line) cells labeling EpCAM with ab213500 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing membranous staining on 4T1 cells.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) 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 (ab213500).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EpCAM antibody [EPR20532-222] - BSA and Azide free (AB232539)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling EpCAM with ab213500 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membranous staining on rat colon is observed [PMID : 15637741].

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213500).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EpCAM antibody [EPR20532-222] - BSA and Azide free (AB232539)

Immunohistochemical analysis of paraffin-embedded human endometrium cancer tissue labeling EpCAM with ab213500 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membranous staining on tumor cells of human endometrium cancer is observed [PMID : 15637741].

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213500).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-EpCAM antibody [EPR20532-222] - BSA and Azide free (AB232539)

Immunofluorescent analysis of 100% methanol-fixed HCT 116 (Human colorectal carcinoma cell line) cells labeling EpCAM with ab213500 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing membranous staining on HCT 116 cells.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) 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 (ab213500).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EpCAM antibody [EPR20532-222] - BSA and Azide free (AB232539)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling EpCAM with ab213500 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membranous staining on human colon is observed [PMID : 15637741].

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213500).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-EpCAM antibody [EPR20532-222] - BSA and Azide free (AB232539)

EpCAM was immunoprecipitated from 0.35 mg of HCT 116 (Human colorectal carcinoma cell line) whole cell lysate with ab213500 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab213500 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : HCT 116 whole cell lysate 10 μg (Input).

Lane 2 : ab213500 IP in HCT 116 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab213500 in HCT 116 whole cell lysate.

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

Exposure time : 30 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213500).

All lanes:

Immunoprecipitation - Anti-EpCAM antibody [EPR20532-222] (<a href='/en-us/products/primary-antibodies/epcam-antibody-epr20532-222-ab213500'>ab213500</a>)

Predicted band size: 35 kDa

false