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  1. Link

    irs2-antibody-epr9042-ab134101.pdf

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Signal Transduction Adapters Cytoplasmic
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-IRS2 antibody [EPR904(2)] (ab134101)

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Western blot - Anti-IRS2 antibody [EPR904(2)] (ab134101)
  • Immunocytochemistry/ Immunofluorescence - Anti-IRS2 antibody [EPR904(2)] (ab134101)
  • Immunocytochemistry/ Immunofluorescence - Anti-IRS2 antibody [EPR904(2)] (ab134101)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
  • Western blot - Anti-IRS2 antibody [EPR904(2)] (ab134101)
  • Western blot - Anti-IRS2 antibody [EPR904(2)] (ab134101)
  • Western blot - Anti-IRS2 antibody [EPR904(2)] (ab134101)
  • Flow Cytometry (Intracellular) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
  • Flow Cytometry (Intracellular) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
  • Anti-IRS2 antibody [EPR904(2)] (ab134101)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR904(2)] to IRS2
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 APC Carrier Free

You may also be interested in

Protein
Product image
Recombinant Human IRS2 protein (ab152902)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Knockout
Product image
Human IRS2 knockout HEK-293 cell line (ab264013)

View more associated products

Overview

  • Product name

    Anti-IRS2 antibody [EPR904(2)]
    See all IRS2 primary antibodies
  • Description

    Rabbit monoclonal [EPR904(2)] to IRS2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human IRS2 aa 1300 to the C-terminus (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HEK293, HEK293 (treated with insulin), and NIH/3T3 (treated with insulin) and SH-SY5Y cell lysates; A375 (human malignant melanoma epithelial cell) and A549 whole cell lysates IHC-P: Human kidney, breast and muscle tissues and rat kidney tissue. ICC/IF: SH-SY5Y cells. HEK293 WT cells (HEK293-IRS2 KO used as a negative cell line). Flow Cyt (intra): HeLa cells.
  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR904(2)
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Adapters
    • Cytoplasmic
    • Signal Transduction
    • Growth Factors/Hormones
    • Insulin / Insulin-like
    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Cardiovascular
    • Atherosclerosis
    • Diabetes associated
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
    • Metabolism
    • Types of disease
    • Diabetes
    • Metabolism
    • Types of disease
    • Heart disease

Associated products

  • Alternative Versions

    • Anti-IRS2 antibody [EPR904(2)] - BSA and Azide free (ab208195)
    • Alexa Fluor® 488 Anti-IRS2 antibody [EPR904(2)] (ab214870)
    • APC Anti-IRS2 antibody [EPR904(2)] (ab221281)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human IRS2 knockout HEK-293 cell line (ab264013)
  • KO cell lysates

    • Human IRS2 knockout HEK-293 cell lysate (ab264506)
  • Positive Controls

    • HEK-293 whole cell lysate (ab7902)
  • Recombinant Protein

    • Recombinant Human IRS2 protein (ab152902)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab134101 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
Flow Cyt (Intra)
1/100 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/1000 - 1/10000. Detects a band of approximately 170-185 kDa (predicted molecular weight: 137 kDa).

Although some papers support the expression in liver (PMID: 30202052), A549 (PMID: 30988063), NCI-H1299 (PMID: 30988063), LADMAC (PMID: 29115630), MDA-MB-231 (PMID: 29685905) and MEF (PMID: 30679431), ab134101 can’t detect the target band in these samples, even at the dilution of 1:200

IHC-P
1/50 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/300.
Notes
Flow Cyt (Intra)
1/100 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/1000 - 1/10000. Detects a band of approximately 170-185 kDa (predicted molecular weight: 137 kDa).

Although some papers support the expression in liver (PMID: 30202052), A549 (PMID: 30988063), NCI-H1299 (PMID: 30988063), LADMAC (PMID: 29115630), MDA-MB-231 (PMID: 29685905) and MEF (PMID: 30679431), ab134101 can’t detect the target band in these samples, even at the dilution of 1:200

IHC-P
1/50 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/300.

Target

  • Function

    May mediate the control of various cellular processes by insulin.
  • Sequence similarities

    Contains 1 IRS-type PTB domain.
    Contains 1 PH domain.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization

    Cytoplasm > cytosol.
  • Target information above from: UniProt accession Q9Y4H2 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 8660 Human
    • Entrez Gene: 384783 Mouse
    • Entrez Gene: 29376 Rat
    • Omim: 600797 Human
    • SwissProt: Q9Y4H2 Human
    • SwissProt: P81122 Mouse
    • Unigene: 442344 Human
    • Unigene: 407207 Mouse
    • Alternative names

      • Insulin receptor substrate 2 antibody
      • IRS 2 antibody
      • IRS-2 antibody
      • Irs2 antibody
      • IRS2_HUMAN antibody
      see all

    Images

    • Western blot - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      Western blot - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      All lanes : Anti-IRS2 antibody [EPR904(2)] (ab134101) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293 cell lysate
      Lane 2 : IRS2 knockout HEK-293 cell lysate
      Lane 3 : A-375 cell lysate
      Lane 4 : A549 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 137 kDa
      Observed band size: 160 kDa why is the actual band size different from the predicted?



      Lanes 1 - 4: Merged signal (red and green). Green - ab134101 observed at 160 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

      ab134101 was shown to react with IRS2 in wild-type HEK-293 cells in western blot with loss of signal observed in IRS2 knockout sample. Wild-type and IRS2 knockout HEK-293 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab134101 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      Immunocytochemistry/ Immunofluorescence - Anti-IRS2 antibody [EPR904(2)] (ab134101)

      ab134101 staining IRS2 in wild-type HEK293 cells (top panel) and IRS2 knockout HEK293 cells (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab134101 at 1/500 and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Immunocytochemistry/ Immunofluorescence - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      Immunocytochemistry/ Immunofluorescence - Anti-IRS2 antibody [EPR904(2)] (ab134101)

      Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y cells labelling IRS2 with purified ab134101 at 1/300. 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 (1/300) 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).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling IRS2 with purified ab134101 at 1/500. Heat mediated antigen retrieval was performed using Tris/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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling IRS2 with purified ab134101 at 1/500. Heat mediated antigen retrieval was performed using Tris/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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human muscle tissue labelling IRS2 with unpurified ab134101 at a dilution of 1/50.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast tissue labelling IRS2 with unpurified ab134101 at a dilution of 1/50.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Western blot - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      Western blot - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      All lanes : Anti-IRS2 antibody [EPR904(2)] (ab134101) at 1/1000 dilution

      Lane 1 : Rat liver lysates
      Lane 2 : Human liver lysates
      Lane 3 : A375 (human malignant melanoma epithelial cell) whole cell lysates
      Lane 4 : A549 (human lung carcinoma epithelial cell) whole cell lysates
      Lane 5 : NCI-H1299 (human lung carcinoma epithelial cell) whole cell lysates
      Lane 6 : LADMAC (Mouse bone marrow monocyte macrophage) whole cell lysates
      Lane 7 : MDA-MB-231 (human breast adenocarcinoma epithelial cell) whole cell lysates
      Lane 8 : MEF (Mouse embryonic fibroblast) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Developed using the ECL technique.

      Predicted band size: 137 kDa
      Observed band size: 170-185 kDa why is the actual band size different from the predicted?



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

      Exposure time:

      Left image: 180 seconds

      Right image: 40 seconds

      Although some papers support the expression in liver (PMID: 30202052), A549 (PMID: 30988063), NCI-H1299 (PMID: 30988063), LADMAC (PMID: 29115630), MDA-MB-231 (PMID: 29685905) and MEF (PMID: 30679431), ab134101 can’t detect the target band in these samples, even at the dilution of 1:200.

    • Western blot - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      Western blot - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      Anti-IRS2 antibody [EPR904(2)] (ab134101) at 1/5000 dilution (purified) + NIH/3T3 whole cell lysate - treated with insulin at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

      Predicted band size: 137 kDa
      Observed band size: 170-185 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      Western blot - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      All lanes : Anti-IRS2 antibody [EPR904(2)] (ab134101) at 20 µg (purified)

      Lane 1 : HEK293 whole cell lysate - untreated
      Lane 2 : HEK293 whole cell lysate - treated with insulin
      Lane 3 : SH-SY5Y whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

      Predicted band size: 137 kDa
      Observed band size: 170-185 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

    • Flow Cytometry (Intracellular) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      Flow Cytometry (Intracellular) - Anti-IRS2 antibody [EPR904(2)] (ab134101)

      Intracellular Flow Cytometry analysis of HeLa cells labelling IRS2 with purified ab134101 at 1/120 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies. 

       

       

    • Flow Cytometry (Intracellular) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
      Flow Cytometry (Intracellular) - Anti-IRS2 antibody [EPR904(2)] (ab134101)

      Overlay histogram showing HeLa cells stained with unpurified ab134101 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab134101, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions. 

       

       

    • Anti-IRS2 antibody [EPR904(2)] (ab134101)
      Anti-IRS2 antibody [EPR904(2)] (ab134101)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (20)

    Publishing research using ab134101? Please let us know so that we can cite the reference in this datasheet.

    ab134101 has been referenced in 20 publications.

    • Chen H & Chen J LncRNA SOX21-AS1 Promotes the Growth and Invasiveness of Osteosarcoma Cells Through miR-7-5p/IRS2 Regulatory Network. Arch Med Res 52:294-303 (2021). PubMed: 33341286
    • Meng L  et al. Pseudo-Starvation Driven Energy Expenditure Negatively Affects Ovarian Follicle Development. Int J Mol Sci 22:N/A (2021). PubMed: 33808081
    • Ettcheto M  et al. Dexibuprofen ameliorates peripheral and central risk factors associated with Alzheimer's disease in metabolically stressed APPswe/PS1dE9 mice. Cell Biosci 11:141 (2021). PubMed: 34294142
    • Yun C  et al. Pilose antler peptide promotes osteoblast proliferation, differentiation and mineralization via the insulin signaling pathway. Exp Ther Med 19:923-930 (2020). PubMed: 32010253
    • Han L  et al. A Novel Antidiabetic Monomers Combination Alleviates Insulin Resistance Through Bacteria-Cometabolism-Inflammation Responses. Front Microbiol 11:173 (2020). PubMed: 32132984
    View all Publications for this product

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    Please use the links above to contact us or submit feedback about this product.

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