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.

ab111912, at a 1/50 dilution, staining Somatostatin 28 in paraffin embedded Human pancreas tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Ab111912 staining Somatostatin-28 in Rat hippocampus tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with 4% paraformaldehyde and permeabilized with 0.2% TritonX-100. Heat mediated antigen retrieval was performed using a citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20).  Primary antibody was used at a 1:500 dilution. Ab150077 (1:1000) was used as the secondary antibody. Counterstained with DAPI. Cytoplasmic staining in the hilus of the dentate gyrus of rat hippocampus [PMID: 25469233] . 

ab111912 staining Somatostatin 28 in Mouse pancreas tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Heat mediated antigen retrieval in a TRIS/EDTA buffer pH9 (ab93684). Primary antibody at 1:4000 dilution and   a  Goat Anti- rabbit IgG H&L (HRP) was used as the secondary antibody. Couterstained with hematoxylin. Cytoplasmic staining on delta cells of mouse pancreas islet [PMID: 25019573], [PMID: 9066918], [BiologiaSerbica 2014, Vol. 36  No. 1-2 24-32] . 

Ab111912 staining Somatostatin-28 in Rat pancreas tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with 4% paraformaldehyde and permeabilized with 0.2% TritonX-100. Heat mediated antigen retrieval was performed using a citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20).Primary antibody was used at a 1:500 dilution. Ab150077 (1:1000) was used as the secondary antibody.Counterstained with DAPI. Cytoplasmic staining on delta cells of rat pancreas islet. [PMID: 25019573], [PMID: 9066918], [BiologiaSerbica 2014, Vol. 36  No. 1-2 24-32] .  

Ab111912 staining Somatostatin-28 in Mouse hippocampus tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with 4% paraformaldehyde and permeabilized with 0.2% TritonX-100.Heat mediated antigen retrieval was performed using a citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20). Primary antibody was used at a 1:500 dilution. Ab150077 (1:1000) was used as the secondary antibody.Counterstained with DAPI.  Cytoplasmic staining in the hilus of the dentate gyrus of mouse hippocampus [PMID: 25469233]. 

Ab111912 staining Somatostatin-28 in Mouse pancreas tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with 4% paraformaldehyde and permeabilized with 0.2% TritonX-100. Heat mediated antigen retrieval was performed using a citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20). Primary antibody was used at a 1:500 dilution. Ab150077 (1:1000) was used as the secondary antibody. Counterstained with DAPI. Cytoplasmic staining on delta cells of mouse pancreas islet. [PMID: 25019573], [PMID: 9066918], [BiologiaSerbica 2014, Vol. 36  No. 1-2 24-32] . 

ab111912 staining Somatostatin 28 in Rat pancreas tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Antigen retrieval was by heat mediated in a TRIS/EDTA buffer pH9 (ab93684).Primary antibody at 1:4000 dilution and Goat Anti- rabbit IgG H&L (HRP) was used as the secondary antibody. Counterstained with Hematoxylin. Cytoplasmic staining on delta cells of rat pancreas islet[PMID: 25019573], [PMID: 9066918], [BiologiaSerbica 2014, Vol. 36  No. 1-2 24-32]  .

ab111912 staining Somatostatin 28 in human pancreas tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Heat mediated antigen retrieval in a TRIS/EDTA buffer pH9 (ab93684). Primary antibody  at 1/4000 dilution and a Goat Anti- rabbit IgG H&L (HRP) was used as the secondary antibody. Counterstained with Hematoxylin. Cytoplasmic staining on delta cells of human pancreas islet. [PMID: 25019573], [PMID: 9066918],  [BiologiaSerbica 2014, Vol. 36  No. 1-2 24-32] .