• Product name
    Anti-IRS2 (phospho S731) antibody
    See all IRS2 primary antibodies
  • Description
    Rabbit polyclonal to IRS2 (phospho S731)
  • Host species
  • Specificity
    This antibody is specific for human IRS2 phosphorylated at the position of Serine 731. The antibodies were evaluated for specificity with a dot blot assay using synthetically prepared IRS peptides. It only recognizes the phosphorylated serine 731, not other phosphorylated sites or non-phosphorylated IRS 1 or IRS 2.
  • Tested applications
    Suitable for: WB, IP, ELISA, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (YKASSpPAESS), corresponding to amino acids 727-736 of human IRS2 at the phosphorylated site of Serine 731. This synthetic peptide also correlates to amino acids 719-728 of mouse IRS2 at the phosphorylation site of Serine 723 and to amino acids 229-238 of rat IRS2 at the serine phosphorylation site 233.



Our Abpromise guarantee covers the use of ab3690 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
WB Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 200 kDa. in cell lysate derived from mouse serum-treated fibroblasts, which corresponds to the predicted molecular weight of phospho IRS 2. Samples were boiled prior to subject 7.5% SDS-polyacrylamide gelelectrophoresis.
IP Use at an assay dependent concentration. Use at a concentration of 3.0 - 5.0 µg / extract from 107 cells.
ELISA Use a concentration of 0.1 - 1 µg/ml.
IHC-P Use a concentration of 2 µg/ml.



  • Anti-IRS2 (phospho S731) antibody (ab3690) at 1 µg/ml + Brain (Mouse) Tissue Lysate at 10 µg

    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 160 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 121 kDa, 64 kDa. We are unsure as to the identity of these extra bands.

    Exposure time: 8 minutes

    IRS2 contains a number of potential phosphorylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. The 160 kDa band observed is also comparable to the molecular weight seen with other commercially available antibodies to IRS2.
  • IP: 3T3 cell lysate immunoprecipitated with Anti-IRS-2 (pSER731) (ab3690)
  • ab3690 (2µg/ml) staining IRS2 (phospho S731) in human cerebellum using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of the cell membrane.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.


This product has been referenced in:
  • Gao W  et al. NEFA-induced ROS impaired insulin signalling through the JNK and p38MAPK pathways in non-alcoholic steatohepatitis. J Cell Mol Med 22:3408-3422 (2018). Read more (PubMed: 29602237) »
  • Jia Y  et al. Dapagliflozin Aggravates Renal Injury via Promoting Gluconeogenesis in db/db Mice. Cell Physiol Biochem 45:1747-1758 (2018). Read more (PubMed: 29495021) »
See all 9 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (7% SDS-PAGE)
Rat Tissue lysate - whole (Muscle, Fat, Liver)
Muscle, Fat, Liver
2U insulin for 5mins
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Feb 18 2013


Thank you for your reply. I apologize for the confusion, but I figured out what the problem was. The BLAST was picking up a fragment of the protein, where that sequence was within the amino acid range I mentioned. In the full length sequence, you are correct that the sequence is found surrounding S723. I have changed the information on our datasheet. I am very sorry about that. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Thank you for your enquiry. I'm not sure I understand your question. On the datasheet, it states that the immunogen peptide sequence "also correlates to amino acids 129-138 of mouse IRS2 at the phosphorylation site of Serine 133 and to amino acids 229-238 of rat IRS2 at the serine phosphorylation site 233". When I BLAST the sequence, I get the same information: Mouse Score = 32.5 bits (69), Expect = 1.4 Identities = 10/10 (100%), Positives = 10/10 (100%)Query: 1 YKASSPAESS 10 YKASSPAESS Sbjct: 129 YKASSPAESS 138 Rat Score = 32.5 bits (69), Expect = 1.4 Identities = 10/10 (100%), Positives = 10/10 (100%) Query: 1 YKASSPAESS 10 YKASSPAESS Sbjct: 229 YKASSPAESS 238 It doesn't appear that this antibody would detect IRS1, as I did not get any results for IRS1 in a BLAST of the immunogen sequence. I also did an alignment of the IRS1 sequence with the immunogen sequence, and did not get any significant homology. What publication was this listed in? Was it this particular antibody? I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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