Anti-Insulin Receptor alpha antibody [EPR23962-157] - BSA and Azide free (ab283717)

Rabbit Recombinant Monoclonal Insulin Receptor antibody. Carrier free. Suitable for IP, WB, ICC/IF, IHC-P and reacts with Human, Mouse, Rat samples.

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Images

Western blot - Anti-Insulin Receptor alpha antibody [EPR23962-157] - BSA and Azide free (AB283717), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	Flow Cyt	WB	ICC/IF	IHC-P
Human	Tested	Not recommended	Tested	Tested	Tested
Mouse	Not recommended	Not recommended	Tested	Not recommended	Tested
Rat	Not recommended	Not recommended	Tested	Expected	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse, Rat, Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target data

See full target information INSR

Function

Receptor tyrosine kinase which mediates the pleiotropic actions of insulin. Binding of insulin leads to phosphorylation of several intracellular substrates, including, insulin receptor substrates (IRS1, 2, 3, 4), SHC, GAB1, CBL and other signaling intermediates. Each of these phosphorylated proteins serve as docking proteins for other signaling proteins that contain Src-homology-2 domains (SH2 domain) that specifically recognize different phosphotyrosine residues, including the p85 regulatory subunit of PI3K and SHP2. Phosphorylation of IRSs proteins lead to the activation of two main signaling pathways: the PI3K-AKT/PKB pathway, which is responsible for most of the metabolic actions of insulin, and the Ras-MAPK pathway, which regulates expression of some genes and cooperates with the PI3K pathway to control cell growth and differentiation. Binding of the SH2 domains of PI3K to phosphotyrosines on IRS1 leads to the activation of PI3K and the generation of phosphatidylinositol-(3, 4, 5)-triphosphate (PIP3), a lipid second messenger, which activates several PIP3-dependent serine/threonine kinases, such as PDPK1 and subsequently AKT/PKB. The net effect of this pathway is to produce a translocation of the glucose transporter SLC2A4/GLUT4 from cytoplasmic vesicles to the cell membrane to facilitate glucose transport. Moreover, upon insulin stimulation, activated AKT/PKB is responsible for: anti-apoptotic effect of insulin by inducing phosphorylation of BAD; regulates the expression of gluconeogenic and lipogenic enzymes by controlling the activity of the winged helix or forkhead (FOX) class of transcription factors. Another pathway regulated by PI3K-AKT/PKB activation is mTORC1 signaling pathway which regulates cell growth and metabolism and integrates signals from insulin. AKT mediates insulin-stimulated protein synthesis by phosphorylating TSC2 thereby activating mTORC1 pathway. The Ras/RAF/MAP2K/MAPK pathway is mainly involved in mediating cell growth, survival and cellular differentiation of insulin. Phosphorylated IRS1 recruits GRB2/SOS complex, which triggers the activation of the Ras/RAF/MAP2K/MAPK pathway. In addition to binding insulin, the insulin receptor can bind insulin-like growth factors (IGFI and IGFII). Isoform Short has a higher affinity for IGFII binding. When present in a hybrid receptor with IGF1R, binds IGF1. PubMed:12138094 shows that hybrid receptors composed of IGF1R and INSR isoform Long are activated with a high affinity by IGF1, with low affinity by IGF2 and not significantly activated by insulin, and that hybrid receptors composed of IGF1R and INSR isoform Short are activated by IGF1, IGF2 and insulin. In contrast, PubMed:16831875 shows that hybrid receptors composed of IGF1R and INSR isoform Long and hybrid receptors composed of IGF1R and INSR isoform Short have similar binding characteristics, both bind IGF1 and have a low affinity for insulin. In adipocytes, inhibits lipolysis (By similarity).

Alternative names

CD220, Insulin receptor, IR

Recommended products

Rabbit Recombinant Monoclonal Insulin Receptor antibody. Carrier free. Suitable for IP, WB, ICC/IF, IHC-P and reacts with Human, Mouse, Rat samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR23962-157
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C

Notes

ab283717 is the carrier-free version of Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The Insulin Receptor alpha (INSTA) sometimes called Insulin Receptor Subunit alpha is an integral component of the insulin receptor complex. It is an important site for insulin binding and exhibits a molecular weight (MW) of approximately 157 kDa. The receptor commonly distributes on the surface of cells with high expression in muscle cells liver cells and adipose tissue. Insulin receptor antibodies can target this protein for various studies including Western blot analyses. The protein structure itself comprises two alpha subunits alongside two beta subunits which together facilitate insulin's physiological effects.

Biological function summary

INSTA plays a critical role in glucose uptake and regulation of energy metabolism. It is an extracellular receptor involved in the formation of a tetrameric complex when interacting with insulin triggering downstream signaling cascades. The receptor helps mediate processes including glycogen synthesis lipid metabolism and protein synthesis through its influence on glucose transport and metabolic enzyme activity. The interaction with insulin generates a conformational change in the receptor which propagates cellular signal transduction essential for metabolic homeostasis.

Pathways

INSTA forms part of the insulin signaling pathway and the PI3K/Akt pathway. These pathways are essential for cellular glucose uptake and ensure energy balance within the body. Insulin receptor interacts with proteins like IRS-1 and PI3K which play roles in mediating insulin's effects. As part of these pathways INSTA assists in the convergence of signals that regulate anabolic and catabolic processes within cells making it integral to maintaining normal metabolic functions.

Associated diseases and disorders

INSTA holds a significant connection to conditions such as type 2 diabetes mellitus and insulin resistance. Both linked to misregulation or dysregulation of insulin receptor functions these disorders underline the importance of the insulin receptor in maintaining glucose homeostasis within the body. Moreover INSTA's association with other proteins such as IRS-1 becomes particularly apparent in these disease states as malfunctioning of these proteins can impair the insulin signaling cascade exacerbating metabolic syndrome. Understanding INSTA's role provides insight into therapeutic targets for managing these related conditions.

Product promise

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11 product images

Western blot - Anti-Insulin Receptor alpha antibody [EPR23962-157] - BSA and Azide free (ab283717)
This data was developed using Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Low expression cell line: T-47D (PMID:1617668, 29099049).
The 220 kDa band is the pro-insulin receptor, while the 135 kDa band is the insulin receptor alpha subunit (PMID: 28765322, 6389544).
Exposure time: 136 seconds
All lanes: Western blot - Anti-Insulin Receptor alpha antibody [EPR23962-157] (Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689) at 1/1000 dilution
Lane 1: IM-9 (human multiple myeloma B Lymphoblast), whole cell lysate at 20 µg
Lane 2: T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 156 kDa
Observed band size: 135 kDa
Western blot - Anti-Insulin Receptor alpha antibody [EPR23962-157] - BSA and Azide free (ab283717)
This data was developed using Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The 220 kDa band is the pro-insulin receptor, while the 135 kDa band is the insulin receptor alpha subunit (PMID: 28765322, 6389544).
Exposure time: 114 seconds
All lanes: Western blot - Anti-Insulin Receptor alpha antibody [EPR23962-157] (Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689) at 1/1000 dilution
Lane 1: Hepa1-6 (mouse hepatoma epithelial cell), whole cell lysate at 20 µg
Lane 2: C2C12 (mouse myoblasts myoblast), whole cell lysate at 20 µg
Lane 3: Mouse liver tissue lysate at 20 µg
Lane 4: AR42J (rat pancreatic tumor epithelial cell), whole cell lysate at 20 µg
Lane 5: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg
Lane 6: HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Predicted band size: 156 kDa
Observed band size: 220 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin Receptor alpha antibody [EPR23962-157] - BSA and Azide free (ab283717)
This data was developed using Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling Insulin Receptor alpha with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 at 1/2000 (1.08 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human liver. The section was incubated with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin Receptor alpha antibody [EPR23962-157] - BSA and Azide free (ab283717)
This data was developed using Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human renal carcinoma tissue labelling Insulin Receptor alpha with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 at 1/2000 (1.08 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining mainly on blood vessels of human renal carcinoma (PMID: 20182859). The section was incubated with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin Receptor alpha antibody [EPR23962-157] - BSA and Azide free (ab283717)
This data was developed using Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labelling Insulin Receptor alpha with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 at 1/2000 (1.08 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human breast cancer (PMID: 17221153). The section was incubated with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin Receptor alpha antibody [EPR23962-157] - BSA and Azide free (ab283717)
This data was developed using Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labelling Insulin Receptor alpha with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 at 1/2000 (0.54 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse liver (PMID: 2688753). The section was incubated with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin Receptor alpha antibody [EPR23962-157] - BSA and Azide free (ab283717)
This data was developed using Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labelling Insulin Receptor alpha with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 at 1/2000 (0.54 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse colon. The section was incubated with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunoprecipitation - Anti-Insulin Receptor alpha antibody [EPR23962-157] - BSA and Azide free (ab283717)
This data was developed using Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689, the same antibody clone in a different buffer formulation.
Insulin Receptor alpha was immunoprecipitated from 0.35 mg IM-9 (human multiple myeloma b lymphoblast) whole cell lysate with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: IM-9 (human multiple myeloma b lymphoblast) whole cell lysate 10 ug
Lane 2: ab2833689 IP in IM-9 whole cell lysate 10 ug
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 in IM-9 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 128 second.
All lanes: Immunoprecipitation - Anti-Insulin Receptor alpha antibody [EPR23962-157] (Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689)
Predicted band size: 156 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin Receptor alpha antibody [EPR23962-157] - BSA and Azide free (ab283717)
This data was developed using Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat liver tissue labelling Insulin Receptor alpha with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 at 1/2000 (0.54 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat liver. The section was incubated with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin Receptor alpha antibody [EPR23962-157] - BSA and Azide free (ab283717)
This data was developed using Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Rat testis tissue labelling Insulin Receptor alpha with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 at 1/2000 (0.54 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on leydig cells of rat testis. The section was incubated with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunocytochemistry/ Immunofluorescence - Anti-Insulin Receptor alpha antibody [EPR23962-157] - BSA and Azide free (ab283717)
This data was developed using Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized IM-9 cells labelling Insulin Receptor alpha with Anti-Insulin Receptor alpha antibody [EPR23962-157] ab283689 at 1/50 (10.8 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing membrane staining in IM-9 cells. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.