Anti-Mast cell protease 1 antibody [EPR25920A-86]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mast cell protease 1 antibody [EPR25920A-86] (AB324633)

Immunohistochemical analysis of formalin fixed paraffin embedded mouse small intestine labelling Mast cell protease 1 with ab324633 at a concentration of 10µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a ChromoMap DAB kit, anti-rabbit HQ and anti-HQ HRP detection. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab324633 anti-Mast cell protease 1 antibody [EPR25920A-86] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

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-Mast cell protease 1 antibody [EPR25920A-86] (AB324633)

Immunohistochemical analysis of paraffin-embedded Mouse brain tissue labeling Mast cell protease 1 with ab324633 at 1/500 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in mouse brain.
The section was incubated with ab324633 for 15 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mast cell protease 1 antibody [EPR25920A-86] (AB324633)

Immunohistochemical analysis of paraffin-embedded Mouse small intestine tissue labeling Mast cell protease 1 with ab324633 at 1/500 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse small intestine.
The section was incubated with ab324633 for 15 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Mast cell protease 1 antibody [EPR25920A-86] (AB324633)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized mouse bone marrow cells (BM), mouse bone marrow-derived mast cells (BMMC) and mouse bone marrow-derived intestinal mucosal mast cells (BMimMC) labelling Mast cell protease 1 with ab324633 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

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

• WB

Lab

Western blot - Anti-Mast cell protease 1 antibody [EPR25920A-86] (AB324633)

Western blot : Anti-Mast cell protease 1 antibody ab324633 staining at 1 µg/mL, shown in green; Mouse anti-Beta Actin ab8226 loading control staining at 1/1000 dilution, shown in magenta. A band was observed at 25-37 kDa. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Mast cell protease 1 antibody [EPR25920A-86] (ab324633) at 1 µg/mL

Lane 1:

Mouse Bone Marrow-derived Mast Cells at 20 µg

Lane 2:

Mouse Intestinal Mucosa-derived Mast Cells at 20 µg

Lane 3:

Mouse brain at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 27 kDa

Observed band size: 25-37 kDa,42 kDa

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