Anti-Insulin antibody [EPR17359] - BSA and Azide free

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Insulin antibody [EPR17359] - BSA and Azide free (AB202760)

Fluorescence multiplex immunohistochemical analysis of the human pancreas (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-Carboxypeptidase A (ab278044, magenta; Opal™690), anti-Cytokeratin 19 (ab195872, green; Opal™520) and anti-Insulin (ab181547, red; Opal™570) on human pancreas. Panel B : anti-Carboxypeptidase A stained on acinar cells. Panel C : anti-Cytokeratin 19 stained on centroacinar cells and ducts. Panel D : anti-Insulin stained on beta cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The section was incubated in three rounds of staining : in the order of ab278044 at 1/4000 dilution (0.135 μg/ml), ab195872 at 1/8000 dilution (0.127 μg/ml), and ab181547 at 1/20000 dilution (0.053 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. DAPI (blue) was used as a nuclear counter stain. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181547).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Insulin antibody [EPR17359] - BSA and Azide free (AB202760)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized BxPC-3 (Human pancreas adenocarcinoma cells) cells labeling Insulin with ab181547 at 1/200 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image shows cytoplasmic staining on BxPC-3 cells. The nuclear counter stain is Dapi (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows : -
-ve control 1 : - ab181547 at 1/200 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : - ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181547).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin antibody [EPR17359] - BSA and Azide free (AB202760)

Immunohistochemical analysis of paraffin-embedded Human adenocarcinoma of colon tissue with ab181547 at 1/64000 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Negative staining on Human colonic adenocarcinoma is observed. Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181547).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin antibody [EPR17359] - BSA and Azide free (AB202760)

Immunohistochemical analysis of paraffin-embedded Human liver tissue with ab181547 at 1/64000 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Negative staining on Human liver tissue is observed. Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181547).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin antibody [EPR17359] - BSA and Azide free (AB202760)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling Insulin with ab181547 at 1/64000 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasm staining on islet cells of Human pancreas is observed. Counter stained with Hematoxylin.

Negative control : PBS instead of primary antibody; secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181547).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Insulin antibody [EPR17359] - BSA and Azide free (AB202760)

This data was developed using ab181547, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse pancreas tissue staining SLC7A5/LAT1 with ab324354 at a 1/2000 dilution, ab278044 anti-Carboxypeptidase A used at 1/4000 dilution and ab181547 anti-Insulin used at a 1/20000 dilution.

Panel A : anti-SLC7A5/LAT1 (green; Opal™520), anti-Carboxypeptidase A (magenta; Opal™690) and anti-Insulin (gray; Opal™570) on mouse pancreas.

Panel B : anti-SLC7A5/LAT1 staining membrane in mouse pancreas.

Panel C : anti-Carboxypeptidase A staining acinar cells in mouse pancreas.

Panel D : anti-Insulin 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 ab324354, ab278044 and ab181547 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-FoFr

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Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Insulin antibody [EPR17359] - BSA and Azide free (AB202760)

Immunohistochemical analysis of 4% paraformaldehyde perfusion fixed, frozen section of Mouse pancreas tissue labeling Insulin with ab181547 at 1/1000 dilution, followed by Donkey anti-rabbit Alexa Fluor 594 at 1/1000 dilution. Cytoplasm staining on islet cells of mouse pancreas is observed. Counter stained with DAPI.

Negative control : PBS instead of primary antibody; secondary antibody is Donkey anti-rabbit Alexa Fluor 594 at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181547).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin antibody [EPR17359] - BSA and Azide free (AB202760)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling Insulin with ab181547 at 1/64000 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasm staining on islet cells of mouse pancreas is observed. Counter stained with Hematoxylin.

Negative control : PBS instead of primary antibody; secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181547).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin antibody [EPR17359] - BSA and Azide free (AB202760)

Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling Insulin with ab181547 at 1/64000 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasm staining on islet cells of rat pancreas is observed. Counter stained with Hematoxylin.

Negative control : PBS instead of primary antibody; secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181547).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.