Anti-TROP2 antibody [EPR20043]

• WB

Lab

Western blot - Anti-TROP2 antibody [EPR20043] (AB214488)

Western blot : Anti-TACSTD2 antibody [EPR20043] (ab214488) staining at 1/2000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab214488 was shown to bind specifically to TACSTD2. A band was observed at 36-70 kDa in wild-type MCF7 cell lysates with no signal observed at this size in TACSTD2 knockout cell line. To generate this image, wild-type and TACSTD2 knockout MCF7 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-TROP2 antibody [EPR20043] (ab214488) at 1/2000 dilution

Lane 1:

Wild-type MCF7 cell lysate at 20 µg

Lane 2:

TACSTD2 knockout MCF7 cell lysate at 20 µg

Lane 3:

HCT 116 cell lysate at 20 µg

Lane 4:

SK-BR-3 cell lysate at 20 µg

Predicted band size: 36 kDa

Observed band size: 36-70 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TROP2 antibody [EPR20043] (AB214488)

Immunohistochemical analysis of formalin fixed paraffin embedded Human skin labelling TROP2 with ab214488 at a concentration of 0.05 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab214488 Anti TROP2 antibody EPR20043 was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining with IgG isotype control antibody ab172730.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TROP2 antibody [EPR20043] (AB214488)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling TROP2 with ab214488 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing membrane and weakly cytoplasmic staining on MCF7 cell line.

The nuclear counterstain is DAPI (blue).

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

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

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TROP2 antibody [EPR20043] (AB214488)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HCT 116 (Human colorectal carcinoma cell line) cells labeling TROP2 with ab214488 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing membrane staining on HCT 116 cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) 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.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-TROP2 antibody [EPR20043] (AB214488)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed MCF7 (Human breast adenocarcinoma cell line) cells labeling TROP2 with ab214488 at 1/60 dilution (red) compared with aRabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody. The permeabilization step was done using 90% methanol for 30 mins at -20 degrees.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TROP2 antibody [EPR20043] (AB214488)

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling TROP2 with ab214488 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Membrane staining on the duct epithelium of human breast 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) at 1/500 dilution.

Perform heat mediated antigen retrieval 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-TROP2 antibody [EPR20043] (AB214488)

Immunohistochemical analysis of paraffin-embedded human cervix cancer tissue labeling TROP2 with ab214488 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Membrane staining on the tumor cells of human cervix cancer 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) at 1/500 dilution.

Perform heat mediated antigen retrieval 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-TROP2 antibody [EPR20043] (AB214488)

Immunohistochemical analysis of paraffin-embedded human skin tissue labeling TROP2 with ab214488 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Membrane staining on the squamous epithelium of human skin 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) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC

AbReview78401****

Immunocytochemistry - Anti-TROP2 antibody [EPR20043] (AB214488)

Immunocytochemistry analysis of acetone-fixed human keratinocytes staining with ab214488 at 1/500 dilution. Secondary antibody was Alexa Fluor® 488 Goat anti-rabbit-IgG at 1/500 dilution. Samples were incubated with the primary antibody with PBS for 1 hour at 21°C. Blocking was done using 4% serum for 30 minutes at 21°C

This image is courtesy of an anonymous Abreview

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TROP2 antibody [EPR20043] (AB214488)

Immunohistochemical analysis of formalin fixed paraffin embedded Human breast labelling TROP2 with ab214488 at a concentration of 0.05 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab214488 Anti TROP2 antibody EPR20043 was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining with IgG isotype control antibody ab172730.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TROP2 antibody [EPR20043] (AB214488)

Immunohistochemical analysis of formalin fixed paraffin embedded wild type (top panel) or knock out (lower panel) MCF7 cells labelling TROP2 with ab214488 at a concentration of 0.03 µg/ml. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with NaCi buffer (pH 6.0, Epitope Retrieval Solution 1) for 20mins.

ab214488 Anti TROP2 antibody EPR20043 was incubated for 15mins at room temperature. Sections were counterstained with Hematoxylin. Images to the left show absence of staining with IgG isotype control antibody ab172730.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TROP2 antibody [EPR20043] (AB214488)

Immunohistochemical analysis of formalin fixed paraffin embedded Human breast adenocarcinoma labelling TROP2 with ab214488 at a concentration of 0.05 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab214488 Anti TROP2 antibody EPR20043 was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining with IgG isotype control antibody ab172730.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TROP2 antibody [EPR20043] (AB214488)

Immunohistochemical analysis of paraffin-embedded mouse skin tissue labeling TROP2 with ab214488 at 1/2000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Membrane staining on the squamous epithelium of mouse skin 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) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TROP2 antibody [EPR20043] (AB214488)

Immunohistochemical analysis of paraffin-embedded rat skin tissue labeling TACD2 with ab214488 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Membrane staining on the squamous epithelium of rat skin 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) at 1/500 dilution.

• WB

Supplier Data

Western blot - Anti-TROP2 antibody [EPR20043] (AB214488)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1/3 : 2 seconds; Lane 2 : 1 minute.

TROP2 is highly glycosylated and appears as band around 37-50kDa.The molecular weight observed is consistent with what has been described in the literature (PMID : 23070813)

All lanes:

Western blot - Anti-TROP2 antibody [EPR20043] (ab214488) at 1/2000 dilution

Lane 1:

HCT 116 (Human colorectal carcinoma cell line) whole cell lysate at 10 µg

Lane 2:

PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 10 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: 36 kDa

Observed band size: 37-50 kDa

false

• WB

Supplier Data

Western blot - Anti-TROP2 antibody [EPR20043] (AB214488)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1/3 : 3 seconds; Lane 2 : 5 seconds; Lane 4 : 15 seconds.

Lanes 1 - 3:

Western blot - Anti-TROP2 antibody [EPR20043] (ab214488) at 1/2000 dilution

Lane 4:

Western blot - Anti-TROP2 antibody [EPR20043] (ab214488) at 1/5000 dilution

Lane 1:

Human breast cancer lysate at 10 µg

Lane 2:

Human skin lysate at 10 µg

Lane 3:

Human placenta lysate at 10 µg

Lane 4:

Human prostate cancer lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 36 kDa

Observed band size: 37-50 kDa

false

• WB

Lab

Western blot - Anti-TROP2 antibody [EPR20043] (AB214488)

Blocking and diluting buffer and concentration : 5% NFDM /TBST. ab181602 was used as a GAPDH loading control. Negative sample : A549 (PMID : 22419550). TROP2 is highly glycosylated and appears as band around 37-50kDa.The molecular weight observed is consistent with what has been described in the literature (PMID : 23070813).

All lanes:

Western blot - Anti-TROP2 antibody [EPR20043] (ab214488) at 1/2000 dilution

Lane 1:

Mouse skin lysate at 20 µg

Lane 2:

Mouse kidney lysate at 20 µg

Lane 3:

Mouse lung lysate at 20 µg

Lane 4:

Rat skin lysate at 20 µg

Lane 5:

Rat lung 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

Predicted band size: 36 kDa

false

Exposure time: 60s