Recombinant Anti-IRS1+IRS2 antibody [EP263Y] (ab40777)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP263Y] to IRS1 + IRS2
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-IRS1+IRS2 antibody [EP263Y]
See all IRS1+IRS2 primary antibodies -
Description
Rabbit monoclonal [EP263Y] to IRS1 + IRS2 -
Host species
Rabbit -
Specificity
The immunogen used for this product shares 78% homology with IRS2. Cross-reactivity with this protein has not been confirmed experimentally.
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Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IF, IHC-Pmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human IRS1 (C terminal). The exact sequence is proprietary.
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Positive control
- WB: HeLa and HEK293 cell lysates, HEK-293T transfected with a human IRS1 expression vector containing a Myc/DDK tag whole cell lysate, HEK-293T transfected with a human IRS2 expression vector containing a Myc/DDK tag whole cell lysate. IHC-P: Human breast carcinoma and cervical carcinoma tissues. ICC/IF: MCF-7 cells.
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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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP263Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab40777 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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB |
1/1000 - 1/10000. Detects a band of approximately 170 kDa (predicted molecular weight: 130 kDa).
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ICC/IF |
1/250 - 1/500.
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
1/1000 - 1/10000. Detects a band of approximately 170 kDa (predicted molecular weight: 130 kDa). |
ICC/IF
1/250 - 1/500. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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Cellular localization
IRS2: Cytoplasm > cytosol. -
Database links
- Entrez Gene: 3667 Human
- Entrez Gene: 8660 Human
- Omim: 147545 Human
- Omim: 600797 Human
- SwissProt: P35568 Human
- SwissProt: Q9Y4H2 Human
- Unigene: 442344 Human
- Unigene: 471508 Human
Images
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Lanes 1-3 : Anti-IRS1+IRS2 antibody [EP263Y] (ab40777) at 1/1000 dilution
Lanes 4-6 : Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [EPR20018-251] (ab205606) at 1/1000 dilution
Lanes 1 & 4 : HEK-293T transfected with an empty vector whole cell lysate
Lanes 2 & 5 : HEK-293T transfected with a human IRS1 expression vector containing a Myc/DDK tag whole cell lysate
Lanes 3 & 6 : HEK-293T transfected with a human IRS2 expression vector containing a Myc/DDK tag whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 130 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM /TBST.
Exposure time: Lane 1, 2, 4, 5 , 6: 1 second. Lane 3: 80 seconds.
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Anti-IRS1+IRS2 antibody [EP263Y] (ab40777) at 1/10000 dilution (purified) + HEK293 whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 130 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?
Blocking and dilution buffer: 5% NFDM/TBST -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labelling IRS1 with purified ab40777 at a dilution of 1/100. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling IRS1 with purified ab40777 at a dilution of 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/250) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
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Anti-IRS1+IRS2 antibody [EP263Y] (ab40777) (purified) + HeLa whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 130 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?
Blocking and dilution buffer: 5% NFDM/TBST -
Intracellular Flow Cytometry analysis of MCF-7 (human breast carcinoma) cells labeling IRS1 with unpurified ab40777 at 1/90 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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Anti-IRS1+IRS2 antibody [EP263Y] (ab40777) at 1/1000 dilution (unpurified) + HeLa cell lysate at 10 µg
Predicted band size: 130 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma labelling IRS1 with unpurified ab40777.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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
References (20)
ab40777 has been referenced in 20 publications.
- Liu X et al. β-Carotene regulates glucose transport and insulin resistance in gestational diabetes mellitus by increasing the expression of SHBG. Clin Exp Pharmacol Physiol 49:1307-1318 (2022). PubMed: 35986631
- Pan S et al. STIM1 promotes angiogenesis by reducing exosomal miR-145 in breast cancer MDA-MB-231 cells. Cell Death Dis 12:38 (2021). PubMed: 33414420
- Zhao L et al. TNF-a promotes insulin resistance in obstructive sleep apnea-hypopnea syndrome. Exp Ther Med 21:568 (2021). PubMed: 33850540
- Su C et al. Roux-en-Y Gastric Bypass Improves Hepatic Glucose Metabolism Involving Upregulation of Sirt1 in Type 2 Diabetes Mellitus. Diabetes Metab Syndr Obes 14:2269-2280 (2021). PubMed: 34045876
- Eyler CE et al. Single-cell lineage analysis reveals genetic and epigenetic interplay in glioblastoma drug resistance. Genome Biol 21:174 (2020). PubMed: 32669109