Anti-Insulin 抗体 [EPR17359]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Insulin antibody [EPR17359] (AB181547)

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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin antibody [EPR17359] (AB181547)

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. Counterstained with hematoxylin.

Perform heat mediated antigen retrieval 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] (AB181547)

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. Counterstained with hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Insulin antibody [EPR17359] (AB181547)

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 counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (Alexa Fluor^® 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 (Alexa Fluor^® 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin antibody [EPR17359] (AB181547)

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. Counterstained with hematoxylin.

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

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Insulin antibody [EPR17359] (AB181547)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse pancreas tissue staining CFTR with ab325171 at a 1 : 2000 (0.259 ug/ml) dilution, ab323181 anti-TMEM16A used at 1 : 1000 (0.532 ug/ml) dilution and ab181547 anti-Insulin used at a 1 : 20000 (0.053 ug/ml) dilution.

Panel A : anti-CFTR (green; Opal™520), anti-TMEM16A (magenta; Opal™690) and anti-Insulin (gray; Opal™570) on mouse pancreas.
Panel B : anti-CFTR staining centroacinar cells in mouse pancreas.
Panel C : anti-TMEM16A staining epithelium 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 ab325171, ab323181 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.

Nuclear counter stain with DAPI.

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] (AB181547)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin antibody [EPR17359] (AB181547)

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. Counterstained with hematoxylin.

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

Perform heat mediated antigen retrieval 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] (AB181547)

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. Counterstained with hematoxylin.

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

Perform heat mediated antigen retrieval 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] (AB181547)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse pancreas tissue staining MafA with ab324869 at a 1 : 100 (5.13 ug/ml) dilution, ab181547 anti-Insulin used at 1 : 20000 (0.053 ug/ml) dilution and ab209792 anti-GIP used at a 1 : 2000 (0.26 ug/ml) dilution.

Panel A : merged staining of anti-MafA (green; Opal™520), anti-Insulin (magenta; Opal™690) and anti-GIP (gray; Opal™570) on mouse pancreas.
Panel B : anti-MafA staining nucleus of beta cells in mouse pancreas islet.
Panel C : anti-Insulin staining cytoplasm of beta cells in mouse pancreas islet.
Panel D : anti-GIP staining the alpha cells in mouse pancreas islet.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324869, ab181547 and ab209792 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin antibody [EPR17359] (AB181547)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat pancreas tissue staining MafA with ab324869 at a 1 : 100 (5.13 ug/ml) dilution, ab181547 anti-Insulin used at 1 : 20000 (0.053 ug/ml) dilution and ab209792 anti-GIP used at a 1 : 2000 (0.26 ug/ml) dilution.

Panel A : merged staining of anti-MafA (green; Opal™520), anti-Insulin (magenta; Opal™690) and anti-GIP (gray; Opal™570) on rat pancreas.
Panel B : anti-MafA staining nucleus of beta cells in rat pancreas islet.
Panel C : anti-Insulin staining cytoplasm of beta cells in rat pancreas islet.
Panel D : anti-GIP staining the alpha cells in rat pancreas islet.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324869, ab181547 and ab209792 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.

• WB

Unknown

Western blot - Anti-Insulin antibody [EPR17359] (AB181547)

Blocking/Diluting buffer and concentration : 5% NFDM/TBST

Observed MW : 12KDa

All lanes:

Western blot - Anti-Insulin antibody [EPR17359] (ab181547) at 1/1000 dilution

Lane 1:

Human pancreas at 20 µg

Lane 2:

Mouse pancreas at 20 µg

Lane 3:

Rat pancreas at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 2000 µg/mL

Predicted band size: 12 kDa

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• WB

Lab

Western blot - Anti-Insulin antibody [EPR17359] (AB181547)

All lanes:

Western blot - Anti-Insulin antibody [EPR17359] (ab181547) at 1/1000 dilution

All lanes:

Mouse pancreas lysate 20 µg at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 12 kDa

Observed band size: 12 kDa

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• WB

CiteAb

Western blot - Anti-Insulin antibody [EPR17359] (AB181547)

Insulin western blot using anti-Insulin antibody [EPR17359] ab181547. Publication image and figure legend from Spohrer, S., Gross, R., et al., 2017, Sci Rep, PubMed 29180680.

ab181547 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab181547 please see the product overview.

Effect of CK2 inhibition on insulin expression and secretion of isolated pancreatic islets. (a) Isolated islets were treated with CX-4945 (10 μM) or DMSO as a solvent control for 24 h. Subsequently, protein extracts were generated and CK2 kinase activity was determined. DMSO treated islets were set 100%. Statistical analysis was performed by using Students t-test, **p < 0.01. (b) Light microscopic images of isolated islets which were treated with CX-4945 (10 μM) or DMSO as a solvent control for 24 h. Treatment with 100 μM H2O2 served as positive control for a treatment with an impact on cell viability. After treatment, islets viability was analysed after trypan blue and neutral red staining with a LEICA DMIL microscope and a LEICA DFC450 C camera. Scale bars : 100 μm. (c) Isolated islets were treated with CX-4945 (10 μM) or DMSO as a solvent control for 24 h and the expression of insulin and β-actin was analysed by Western blot. Full-length blots are presented in Supplementary Figure S2. (d) Quantitative analysis of (c). DMSO-treated islets were used as control and set 100%. Statistical analysis was performed by using Students t-test, *p < 0.05. (e) Pancreatic islets were treated with DMSO or 10 μM CX-4945 for 18 hours. After a glucose stimulus secreted insulin was determined in the cell culture supernatant with the rat/mouse insulin ELISA kit. DMSO treated islets were set 100%. Statistical analysis was performed by using Students t-test, **p < 0.01.

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