Anti-EMP-1 antibody [EPR30304-517]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EMP-1 antibody [EPR30304-517] (AB325387)

Immunohistochemical analysis of paraffin-embedded Human skin tissue labeling EMP-1 with ab325387 at 1/100 (4.96 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human skin. The primary antibody was incubated 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

Lab

Immunocytochemistry/ Immunofluorescence - Anti-EMP-1 antibody [EPR30304-517] (AB325387)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized U-87 MG (human glioblastoma-astrocytoma epithelial cell) labelling EMP-1 with ab325387 at 1/200 (2.48 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing membrane and cytoplasmic staining in U-87 MG 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).
Low expression : K-562 (human chronic myelogenous leukemia lymphoblast).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab325387 at 1 : 200 dilution, followed by ab150120 at 1 : 1000 dilution
-ve control 2 : ab7291 at 1 : 1000 dilution, followed by ab150081 at 1 : 1000 dilution

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-EMP-1 antibody [EPR30304-517] (AB325387)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized BxPC-3 (human pancreas adenocarcinoma epithelial cell) labelling EMP-1 with ab325387 at 1/200 (2.48 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing membrane and cytoplasmic staining in BxPC-3 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).
Low expression : Daudi (human Burkitt's lymphoma lymphoblast).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab325387 at 1 : 200 dilution, followed by ab150120 at 1 : 1000 dilution
-ve control 2 : ab7291 at 1 : 1000 dilution, followed by ab150081 at 1 : 1000 dilution

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EMP-1 antibody [EPR30304-517] (AB325387)

Immunohistochemical analysis of paraffin-embedded (A) BxPC-3 (human pancreas adenocarcinoma epithelial cell) cell pellet (B) U-87 MG (human glioblastoma-astrocytoma epithelial cell) cell pellet (C) K-562 (human chronic myelogenous leukemia lymphoblast) cell pellet (D) Daudi (human Burkitt's lymphoma lymphoblast) cell pellet tissue labeling EMP-1 with ab325387 at 1/2000 (0.248 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on BxPC-3 (image A) and U-87 MG (image B), no staining on K-562 (image C) and Daudi (image D). The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EMP-1 antibody [EPR30304-517] (AB325387)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling EMP-1 with ab325387 at 1/100 (4.96 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human placenta. The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EMP-1 antibody [EPR30304-517] (AB325387)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling EMP-1 with ab325387 at 1/100 (4.96 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon. The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EMP-1 antibody [EPR30304-517] (AB325387)

Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling EMP-1 with ab325387 at 1/100 (4.96 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human ovarian carcinoma. The primary antibody was incubated 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

Lab

Western blot - Anti-EMP-1 antibody [EPR30304-517] (AB325387)

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

EMP-1 is a glycoprotein of approximately 20-50 kDa and detected as a 15-kDa band after treated with Peptide : N-glycosidase F (PNGase F).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-EMP-1 antibody [EPR30304-517] (ab325387) at 1/1000 dilution

Lane 1:

Untreated U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate, at 20 µg

Lane 2:

U-87 MG whole cell lysate treated with Peptide: N-glycosidase F (PNGase F), at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 20-50 kDa,15 kDa,36 kDa

false

Exposure time: 26s

• WB

Lab

Western blot - Anti-EMP-1 antibody [EPR30304-517] (AB325387)

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

Low expression : K-562, Daudi.

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-EMP-1 antibody [EPR30304-517] (ab325387) at 1/1000 dilution

Lane 1:

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

Lane 2:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 4:

Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 20-50 kDa,36 kDa

false

Exposure time: 37s

• WB

Lab

Western blot - Anti-EMP-1 antibody [EPR30304-517] (AB325387)

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

Low expression : cerebellum.

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-EMP-1 antibody [EPR30304-517] (ab325387) at 1/1000 dilution

Lane 1:

Human colon tissue lysate at 20 µg

Lane 2:

Human esophagus lysate at 20 µg

Lane 3:

Human cerebellum lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 20-25 kDa,36 kDa

false

Exposure time: 37s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EMP-1 antibody [EPR30304-517] (AB325387)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a human EMP1 expression vector containing a Myc-His tag (B) HEK-293T transfected with a human EMP2 expression vector containing a Myc-His tag (C) HEK-293T transfected with empty vector containing a Myc-His tag tissue labeling EMP-1 with ab325387 at 1/2000 (0.248 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on HEK-293T transfected with a human EMP1 expression vector containing a Myc-His tag (image A), no staining on HEK-293T transfected with a human EMP2 expression vector containing a Myc-His tag (image B) and HEK-293T transfected with empty vector containing a Myc-His tag (image D). The primary antibody was incubated 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

Lab

Western blot - Anti-EMP-1 antibody [EPR30304-517] (AB325387)

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

This antibody does not cross-react with human EMP2.

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-His tag® antibody - (ab15149) (1 : 1000)(20-50KDa);Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-EMP-1 antibody [EPR30304-517] (ab325387) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a his-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human EMP1 expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human EMP2 expression vector containing a his-tag, whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 20-50 kDa,36 kDa

false

Exposure time: 1s