Anti-Trypsin antibody [EPR29805-7]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Trypsin antibody [EPR29805-7] (AB325233)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse pancreas tissue staining Robo4 with ab300046 at a 1/100 (3.75 μg/ml) dilution, ab325233 anti-Trypsin used at a 1/500 (0.98 μg/ml) dilution and ab254259 anti-Amylin used at a 1/2000 (0.26 μg/ml) dilution.

Panel A : anti-Robo4 (green; Opal™570), anti-Trypsin (magenta; Opal™690), anti-Amylin (gray; Opal™570) on mouse pancreas.

Panel B : anti-Robo4 staining endothelium in mouse pancreas.

Panel C : anti-Trypsin staining exocrine gland in mouse pancreas.

Panel D : anti-Amylin staining islet cells in mouse pancreas.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab300046, ab325233 and ab254259 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Trypsin antibody [EPR29805-7] (AB325233)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat pancreas tissue staining Trypsin with ab325233 at a 1/500 (0.98 μg/ml) dilution, ab317606 anti-SHANK2 used at a 1/100 (5.21 μg/ml) dilution and ab254259 anti-Amylin used at a 1/2000 (0.26 μg/ml) dilution.

Panel A : anti-Trypsin (green; Opal™690), anti-SHANK2 (magenta; Opal™520), anti-Amylin (gray; Opal™570) on rat pancreas.

Panel B : anti-Trypsin staining exocrine gland in rat pancreas.

Panel C : anti-SHANK2 staining pancreatic ducts in rat pancreas.

Panel D : anti-Amylin staining islet cells in rat pancreas.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325233, ab317606 and ab254259 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Trypsin antibody [EPR29805-7] (AB325233)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat pancreas tissue staining Robo4 with ab300046 at a 1/100 (3.75 μg/ml) dilution, ab325233 anti-Trypsin used at a 1/500 (0.98 μg/ml) dilution and ab254259 anti-Amylin used at a 1/2000 (0.26 μg/ml) dilution.

Panel A : anti-Robo4 (green; Opal™570), anti-Trypsin (magenta; Opal™690), anti-Amylin (gray; Opal™570) on rat pancreas.

Panel B : anti-Robo4 staining endothelium in rat pancreas.

Panel C : anti-Trypsin staining exocrine gland in rat pancreas.

Panel D : anti-Amylin staining islet cells in rat pancreas.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab300046, ab325233 and ab254259 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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-Trypsin antibody [EPR29805-7] (AB325233)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling Trypsin with ab325233 at 1/500 (0.98 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse pancreas. 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Trypsin antibody [EPR29805-7] (AB325233)

Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling Trypsin with ab325233 at 1/500 (0.98 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat pancreas. 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

Supplier Data

Western blot - Anti-Trypsin antibody [EPR29805-7] (AB325233)

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

Negative control : kidney, heart.

Observed bands at 26kda and 28kDa are trypsin and trypsinogen respectively due to autoactivation of trypsinogen during sample preparation (PMID : 23814066).

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

All lanes:

Western blot - Anti-Trypsin antibody [EPR29805-7] (ab325233) at 1/1000 dilution

Lane 1:

Mouse pancreas tissue lysate at 20 µg

Lane 2:

Mouse kidney tissue lysate at 20 µg

Lane 3:

Mouse heart 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: 36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Trypsin antibody [EPR29805-7] (AB325233)

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

Negative control : kidney, heart.

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

All lanes:

Western blot - Anti-Trypsin antibody [EPR29805-7] (ab325233) at 1/1000 dilution

Lane 1:

Rat pancreas tissue lysate at 20 µg

Lane 2:

Rat kidney tissue lysate at 20 µg

Lane 3:

Rat heart 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: 28 kDa,36 kDa

false

Exposure time: 10s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Trypsin antibody [EPR29805-7] (AB325233)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Trypsin with ab325233 at 1/500 (0.98 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse kidney. 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Trypsin antibody [EPR29805-7] (AB325233)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Trypsin with ab325233 at 1/500 (0.98 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat kidney. 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

Supplier Data

Western blot - Anti-Trypsin antibody [EPR29805-7] (AB325233)

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

This antibody cross-reacts with overexpressed mouse PRSS2 and PRSS3 by western blot.

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

All lanes:

Western blot - Anti-Trypsin antibody [EPR29805-7] (ab325233) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 2:

293T transfected with mouse PRSS1 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T transfected with mouse PRSS2 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 4:

293T transfected with mouse PRSS3 expression vector containing a myc-His-tag® 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: 26 kDa,28 kDa,36 kDa

false

Exposure time: 180s