Recombinant Anti-Insulin Receptor beta antibody [EPR23566-103] (ab278100)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23566-103] to Insulin Receptor beta
- Suitable for: IHC-P, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Insulin Receptor beta antibody [EPR23566-103]
See all Insulin Receptor beta primary antibodies -
Description
Rabbit monoclonal [EPR23566-103] to Insulin Receptor beta -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WBmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: MDA-MB-231, PC-3, HepG2, LNCaP and C2C12 whole cell lysate; Human liver and kidney tissue lysate; Mouse liver and skeletal muscle tissue lysate; Rat liver tissue lysate; AR42J whole cell lysate; Rat-1 whole cell lysate; Recombinant mouse INSR active protein (aa28-748 & aa753-946). IHC-P: Human liver tissue. Human kidney cancer tissue; Mouse and rat liver tissue.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23566-103 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab278100 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-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000.
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Notes |
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IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. |
Target
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Relevance
Insulin receptor mediates the biological activities of insulin by regulating multiple signaling pathways through activation of a series of phosphorylation cascades. The human insulin receptor is a heterotetrameric membrane glycoprotein consisting of disulfide-linked subunits in a ß-a-a-ß configuration. The ß-subunit (95kDa) possesses a single transmembrane domain with tyrosine kinase acivity, whereas the a-subunit (135kDa) is completely extracellular. The alpha subunits each contain insulin binding sites and are entirely extracellular in localization. The beta subunits each possess an extracellular domain, a single transmembrane domain, and a cytoplasmic tyrosine kinase domain. Binding of insulin to the alpha subunits induces a conformation change in the receptor which activates the kinase domain, stimulating tyrosine autophosphorylation of the receptor and tyrosine phosphorylation of at least five different insulin receptor substrates designated IRS-1-4, and Shc. -
Cellular localization
Membrane; Single pass type I membrane protein. -
Database links
- Entrez Gene: 3643 Human
- Entrez Gene: 16337 Mouse
- Entrez Gene: 24954 Rat
- Omim: 147670 Human
- SwissProt: P06213 Human
- SwissProt: P15208 Mouse
- SwissProt: P15127 Rat
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Alternative names
- CD220 antibody
- HHF5 antibody
- HIR B antibody
see all
Images
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All lanes : Anti-Insulin Receptor beta antibody [EPR23566-103] (ab278100) at 1/1000 dilution
Lane 1 : T47D (human ductal breast epithelial tumor epithelial cell), whole cell lysate
Lane 2 : MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 3 : PC-3 (human prostate adenocarcinoma epithelial cell), whole cell lysate
Lane 4 : HepG2 (human hepatocellar carcinoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Observed band size: 45,95,210 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Bands of 95KDa and 45KDa respectively represent full length and the proteolytic cleavage fragment (β') of insulin receptor beta subunit.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:19017805, 20655470, 2859121, 1617668, 29099049).
Low expression: T-47D (PMID: 1617668, 29099049)
Exposure time: 3 minutes
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Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Insulin Receptor beta with ab278100 at 1/2000 (0.24 µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human liver. The section was incubated with ab278100 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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All lanes : Anti-Insulin Receptor beta antibody [EPR23566-103] (ab278100) at 1/1000 dilution
Lane 1 : Human liver tissue lysate
Lane 2 : Human kidney tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Observed band size: 45,95 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Bands of 95KDa and 45KDa respectively represent full length and the proteolytic cleavage fragment (β') of insulin receptor beta subunit.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:19017805, 20655470, 2859121, 1617668, 29099049).
Exposure time: 3 minutes
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Immunohistochemical analysis of paraffin-embedded human kidney cancer tissue labeling Insulin Receptor beta with ab278100 at 1/2000 (0.24 µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Mainly positive staining on endothelial cells of human kidney cancer (PMID:20182859). The section was incubated with ab278100 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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All lanes : Anti-Insulin Receptor beta antibody [EPR23566-103] (ab278100) at 1/1000 dilution
Lane 1 : LNCaP (human prostate carcinoma epithelial cell), whole cell lysate
Lane 2 : LNCaP (human prostate carcinoma epithelial cell) treated with PNGase F whole cell lysate
Lane 3 : C2C12 (mouse myoblasts myoblast), whole cell lysate
Lane 4 : C2C12 (mouse myoblasts myoblast) treated with PNGase F whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Observed band size: 45,95,210 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Bands of 95KDa and 45KDa respectively represent full length and the proteolytic cleavage fragment (β') of insulin receptor beta subunit.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:19017805, 20655470, 2859121, 1617668, 29099049).
This blot was developed using a higher sensitivity ECL substrate.
Exposure time: 3 minutes
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Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling Insulin Receptor beta with ab278100 at 1/2000 (0.24 µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse liver. The section was incubated with ab278100 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
All lanes : Anti-Insulin Receptor beta antibody [EPR23566-103] (ab278100) at 1/1000 dilution
Lane 1 : Mouse liver tissue lysate
Lane 2 : Mouse skeletal muscle tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Observed band size: 45,95,210 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Bands of 95KDa and 45KDa respectively represent full length and the proteolytic cleavage fragment (β') of insulin receptor beta subunit.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:19017805, 20655470, 2859121, 1617668, 29099049).
This blot was developed using a higher sensitivity ECL substrate.
Exposure time: 3 minutes
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All lanes : Anti-Insulin Receptor beta antibody [EPR23566-103] (ab278100) at 1/1000 dilution
Lane 1 : Rat liver tissue lysate
Lane 2 : AR42J (rat pancreatic tumor epithelial cell), whole cell lysate
Lane 3 : Rat-1 (rat embryonic fibroblast), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Observed band size: 45,95,210 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Bands of 95KDa and 45KDa respectively represent full length and the proteolytic cleavage fragment (β') of insulin receptor beta subunit.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:19017805, 20655470, 2859121, 1617668, 29099049).
This blot was developed using a higher sensitivity ECL substrate.
Exposure time: 3 minutes
-
Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Insulin Receptor beta with ab278100 at 1/2000 (0.24 µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat liver. The section was incubated with ab278100 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Anti-Insulin Receptor beta antibody [EPR23566-103] (ab278100) at 1/1000 dilution + Recombinant mouse INSR active protein (aa 28-748 & aa 753-946), 20 ng
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Observed band size: 150,82 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
This blot was developed using a higher sensitivity ECL substrate.
Exposure time: 3 minutes
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab278100 has been referenced in 1 publication.
- Park J et al. Activation of the insulin receptor by an insulin mimetic peptide. Nat Commun 13:5594 (2022). PubMed: 36151101