Name: Anti-Insulin antibody [EPR17359]
Brand: RabMAb
SKU: ab181547
Rating: 5 (11 reviews)

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Multiplex immunohistochemistry - Anti-Insulin antibody [EPR17359] (ab181547)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR17359] to Insulin
Suitable for: IHC-FoFr, WB, IHC-P, ICC/IF, mIHC
Reacts with: Mouse, Rat, Human

Carrier Free PE

Product name

Anti-Insulin antibody [EPR17359]
See all Insulin primary antibodies
Description

Rabbit monoclonal [EPR17359] to Insulin
Host species

Rabbit
Specificity

The human recommendation is based on the IHC-P results. We do not guarantee WB for human.
Tested applications

Suitable for: IHC-FoFr, WB, IHC-P, ICC/IF, mIHCmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- IHC-P: Human, mouse and rat pancreas tissue. ICC/IF: BxPC-3 cells. IHC-FoFr: Mouse pancreas tissue. WB: Mouse & Rat pancreas lysate. mIHC: Human pancreas tissue.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR17359
Isotype

IgG
Research areas

Alternative Versions
- Anti-Insulin antibody [EPR17359] - BSA and Azide free (ab202760)
- PE Anti-Insulin antibody [EPR17359] (ab213192)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human Insulin protein (Active) (ab123768)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab181547 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
IHC-FoFr		1/1000.
WB	(1)	Use at an assay dependent concentration. Predicted molecular weight: 12 kDa.
IHC-P	(7)	1/64000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/200.
mIHC		1/20000.

Notes
IHC-FoFr 1/1000.
WB Use at an assay dependent concentration. Predicted molecular weight: 12 kDa.
IHC-P 1/64000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/200.
mIHC 1/20000.

Function

Insulin decreases blood glucose concentration. It increases cell permeability to monosaccharides, amino acids and fatty acids. It accelerates glycolysis, the pentose phosphate cycle, and glycogen synthesis in liver.
Involvement in disease

Defects in INS are the cause of familial hyperproinsulinemia (FHPRI) [MIM:176730].
Defects in INS are a cause of diabetes mellitus insulin-dependent type 2 (IDDM2) [MIM:125852]. IDDM2 is a multifactorial disorder of glucose homeostasis that is characterized by susceptibility to ketoacidosis in the absence of insulin therapy. Clinical fetaures are polydipsia, polyphagia and polyuria which result from hyperglycemia-induced osmotic diuresis and secondary thirst. These derangements result in long-term complications that affect the eyes, kidneys, nerves, and blood vessels.
Defects in INS are a cause of diabetes mellitus permanent neonatal (PNDM) [MIM:606176]. PNDM is a rare form of diabetes distinct from childhood-onset autoimmune diabetes mellitus type 1. It is characterized by insulin-requiring hyperglycemia that is diagnosed within the first months of life. Permanent neonatal diabetes requires lifelong therapy.
Defects in INS are a cause of maturity-onset diabetes of the young type 10 (MODY10) [MIM:613370]. MODY10 is a form of diabetes that is characterized by an autosomal dominant mode of inheritance, onset in childhood or early adulthood (usually before 25 years of age), a primary defect in insulin secretion and frequent insulin-independence at the beginning of the disease.
Sequence similarities

Belongs to the insulin family.
Cellular localization

Secreted.
Target information above from: UniProt accession P01308 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3630 Human
- Entrez Gene: 16333 Mouse
- Entrez Gene: 24505 Rat
- Omim: 176730 Human
- SwissProt: P01308 Human
- SwissProt: P01325 Mouse
- SwissProt: P01322 Rat
- Unigene: 272259 Human
Alternative names
- IDDM antibody
- IDDM1 antibody
- IDDM2 antibody
- ILPR antibody
- ins antibody
- INS_HUMAN antibody
- Insulin A chain antibody
- Insulin B chain antibody
- IRDN antibody
- MODY10 antibody
- Preproinsulin antibody
- Proinsulin antibody
- Proinsulin precursor antibody
see all

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.
Western blot - Anti-Insulin antibody [EPR17359] (ab181547)

All lanes : Anti-Insulin antibody [EPR17359] (ab181547) at 1/1000 dilution

Lane 1 : Human pancreas
Lane 2 : Mouse pancreas
Lane 3 : Rat pancreas

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 2000 µg/ml

Predicted band size: 12 kDa

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

Observed MW: 12KDa
Western blot - Anti-Insulin antibody [EPR17359] (ab181547)

Anti-Insulin antibody [EPR17359] (ab181547) at 1/1000 dilution + Mouse pancreas lysate 20 µg at 20 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 12 kDa
Observed band size: 12 kDa
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.
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.
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.
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.
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.
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.
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.
Anti-Insulin antibody [EPR17359] (ab181547)

Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab181547? Please let us know so that we can cite the reference in this datasheet.

ab181547 has been referenced in 76 publications.

Yan H et al. Intracellular ATP Signaling Contributes to FAM3A-Induced PDX1 Upregulation in Pancreatic Beta Cells. Exp Clin Endocrinol Diabetes 130:498-508 (2022). PubMed: 34592773
Szczerbinska I et al. Large-Scale Functional Genomics Screen to Identify Modulators of Human β-Cell Insulin Secretion. Biomedicines 10:N/A (2022). PubMed: 35052782
Serrano J et al. Saccharin Stimulates Insulin Secretion Dependent on Sweet Taste Receptor-Induced Activation of PLC Signaling Axis. Biomedicines 10:N/A (2022). PubMed: 35052799
Shi YL et al. Trilobatin, a Natural Food Additive, Exerts Anti-Type 2 Diabetes Effect Mediated by Nrf2/ARE and IRS-1/GLUT2 Signaling Pathways. Front Pharmacol 13:828473 (2022). PubMed: 35153796
Zhang X et al. Micro-RNA-124-5p promotes insulin producing cell differentiation through regulating transcriptional factor NKX6.1. Biochem Biophys Rep 30:101273 (2022). PubMed: 35592615

View all Publications for this product

Customer reviews and Q&As

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1-10 of 11 Abreviews or Q&A

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Cat Tissue sections (Pancreas)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrate, pH 6.0

Permeabilization

Yes - 3% H2O2

Specification

Pancreas

Blocking step

Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 20°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Ms. Amanda La Coco

Verified customer

Submitted Aug 27 2021

Application

Western blot

Sample

Cow Recombinant protein (Total protein from transiently transformed plant)

Gel Running Conditions

Reduced Denaturing (12% gel)

Loading amount

0 cells

Specification

Total protein from transiently transformed plant

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 26 2021

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Dog Tissue sections (Pancreatic tissue (left lobe))

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH 6.0

Permeabilization

Yes - 0.25% Triton X-100

Specification

Pancreatic tissue (left lobe)

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Nicola Marrano

Verified customer

Submitted Jan 26 2021

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Dog Tissue sections (Pancreas)

Antigen retrieval step

Enzymatic - Buffer/Enzyme Used: EDTA

Permeabilization

Specification

Pancreas

Blocking step

No blocking step used for 1 hour(s) and 0 minute(s) · Temperature: 24°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Han-Byul Lee

Verified customer

Submitted May 30 2019

Application

Immunocytochemistry

Sample

Human Cultured Cells (Human)

Permeabilization

Yes - Triton-x

Specification

Human

Blocking step

BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 22 2019

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Rat Tissue sections (Zucker rat pancreas)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris/EDTA

Permeabilization

Yes - 3% H2O2

Specification

Zucker rat pancreas

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 25 2019

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (Pancreas)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrate buffer

Permeabilization

Specification

Pancreas

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 31 2018

Application

Immunohistochemistry (Frozen sections)

Sample

Mouse Tissue sections (pancreas)

Permeabilization

Specification

pancreas

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 22 2017

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (pancreas)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrat-Buffer pH 6,0

Permeabilization

Specification

pancreas

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 17 2017

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (Pancreas)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris/EDTA PH 9

Specification

Pancreas

Blocking step

Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10%

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Toloo Taghian

Verified customer

Submitted Apr 22 2016

1-10 of 11 Abreviews or Q&A

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (76)

Customer reviews and Q&As

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

Western blot abreview for Anti-Insulin antibody [EPR17359]

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

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

Immunocytochemistry abreview for Anti-Insulin antibody [EPR17359]

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

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

Immunohistochemistry (Frozen sections) abreview for Anti-Insulin antibody [EPR17359]

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

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