Anti-Pancreatic Polypeptide antibody [EPR22853-61]

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Pancreatic Polypeptide antibody [EPR22853-61] (AB255827)

Multiplex immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat pancreas tissue.

Panel A : Merged staining of anti-Pancreatic Polypeptide (ab255827, green), anti-Somatostatin 28 (ab111912, red) and DAPI (blue).

Panel B : Anti-Pancreatic Polypeptide (green) stained on PP cells in rat pancreas islet.

Panel C : Anti-Somatostatin 28 (red) stained on delta cells in rat pancreas islet.

Panel D : Nuclear counter stain, DAPI (blue).

Key protocol steps : The section was incubated in two rounds of staining with ab255827 (1/20000 dilution) and ab111912 (1/2000 dilution) for 30 mins at room temperature. Each round was followed by tyramide signal amplification with the appropriate fluorophore. Heat mediated antigen retrieval was used (Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins after every round of antibody/fluorophore staining.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

A ready-to-use anti-Rabbit and Mouse Polymer HRP was used as a secondary.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pancreatic Polypeptide antibody [EPR22853-61] (AB255827)

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labeling Pancreatic Polypeptide with ab255827 at 1/5000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on PP cells in mouse pancreas islet (PMID : 27270601) is observed. Counter stained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

The section was incubated with ab255827 for 10 mins at RT.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Pancreatic Polypeptide antibody [EPR22853-61] (AB255827)

Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized mouse pancreas tissue labeling Pancreatic Polypeptide with ab255827 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). ab255827 and ab6995 are shown to stain PP cells and beta cells of mouse pancreatic islet respectively (PMID : 24825414). The nuclear counter stain is DAPI (blue). Insulin is detected using ab6995, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed (ab150120) at 1/1000 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) secondary antibody at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Pancreatic Polypeptide antibody [EPR22853-61] (AB255827)

Immunohistochemical analysis of frozen section of 4%PFA-fixed, 0.2% Triton X-100 permeabilized rat pancreas tissue labeling Pancreatic Polypeptide with ab255827 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). ab255827 and ab6995 are shown to stain PP cells and beta cells of rat pancreatic islets respectively (PMID : 24825414). The nuclear counter stain is DAPI (blue). Insulin is detected using ab6995, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed (ab150120) at 1/1000 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) secondary antibody at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pancreatic Polypeptide antibody [EPR22853-61] (AB255827)

Immunohistochemical analysis of paraffin-embedded rat pancreas tissue labeling Pancreatic Polypeptide with ab255827 at 1/5000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on PP cells in rat pancreas islet (PMID : 27270601) is observed. Counter stained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

The section was incubated with ab255827 for 10 mins at RT.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Pancreatic Polypeptide antibody [EPR22853-61] (AB255827)

Fluorescence multiplex immunohistochemical analysis of the rat pancreas (Formalin/PFA-fixed paraffin-embedded sections).

Panel A : merged staining of anti-Somatostatin 28 (yellow; Opal™520), anti-GIP (magenta; Opal™570) and anti-Pancreatic Polypeptide (green; Opal™690) on rat pancreas.
Panel B : anti-Somatostatin 28 staining the delta cells in rat pancreas islet.
Panel C : anti-GIP staining the alpha cells in rat pancreas islet.
Panel D : anti-Pancreatic Polypeptide staining the PP cells in rat pancreas islet.

Nuclear DNA was labelled with DAPI (shown in blue). The section was incubated in three rounds of staining : in the order of ab111912 at 1/9000 dilution (0.07 μg/ml), ab271989 at 1/4000 dilution (0.25 μg/ml) and ab255827 at 1/10000 dilution (0.05 μg/ml) 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Pancreatic Polypeptide antibody [EPR22853-61] (AB255827)

Fluorescence multiplex immunohistochemical analysis of the rat pancreas (Formalin/PFA-fixed paraffin-embedded sections).

Panel A : merged staining of anti-SV2A (yellow; Opal™520), anti-GIP (magenta; Opal™570) and anti-Pancreatic Polypeptide (green; Opal™690) on rat pancreas.
Panel B : anti-SV2A staining predominantly the beta cells in mouse pancreas islet.
Panel C : anti-GIP staining the alpha cells in rat pancreas islet.
Panel D : anti-Pancreatic Polypeptide staining the PP cells in rat pancreas islet.

Nuclear DNA was labelled with DAPI (shown in blue). The section was incubated in three rounds of staining : in the order of ab254351 at 1/1000 dilution (0.48 μg/ml), ab271989 at 1/4000 dilution (0.25 μg/ml) and ab255827 at 1/10000 dilution (0.05 μg/ml) 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Pancreatic Polypeptide antibody [EPR22853-61] (AB255827)

Fluorescence multiplex immunohistochemical analysis of the mouse pancreas (Formalin/PFA-fixed paraffin-embedded sections).

Panel A : merged staining of anti-SV2A (yellow; Opal™520), anti-GIP (magenta; Opal™570) and anti-Pancreatic Polypeptide (green; Opal™690) on mouse pancreas.
Panel B : anti-SV2A staining predominantly the beta cells in mouse pancreas islet.
Panel C : anti-GIP staining the alpha cells in mouse pancreas islet.
Panel D : anti-Pancreatic Polypeptide staining the PP cells in mouse pancreas islet.

Nuclear DNA was labelled with DAPI (shown in blue). The section was incubated in three rounds of staining : in the order of ab254351 at 1/1000 dilution (0.48 μg/ml), ab271989 at 1/4000 dilution (0.25 μg/ml) and ab255827 at 1/10000 dilution (0.05 μg/ml) 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Pancreatic Polypeptide antibody [EPR22853-61] (AB255827)

Fluorescence multiplex immunohistochemical analysis of the mouse pancreas (Formalin/PFA-fixed paraffin-embedded sections).

Panel A : merged staining of anti-Somatostatin 28 (yellow; Opal™520), anti-GIP (magenta; Opal™570) and anti-Pancreatic Polypeptide (green; Opal™690) on mouse pancreas.
Panel B : anti-Somatostatin 28 staining the delta cells in mouse pancreas islet.
Panel C : anti-GIP staining the alpha cells in mouse pancreas islet.
Panel D : anti-Pancreatic Polypeptide staining the PP cells in mouse pancreas islet.

Nuclear DNA was labelled with DAPI (shown in blue). The section was incubated in three rounds of staining : in the order of ab111912 at 1/9000 dilution (0.07 μg/ml), ab271989 at 1/4000 dilution (0.25 μg/ml) and ab255827 at 1/10000 dilution (0.05 μg/ml) 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.