Overview

  • Product nameAnti-IRS2 (phospho S731) antibody
    See all IRS2 primary antibodies
  • Description
    Rabbit polyclonal to IRS2 (phospho S731)
  • SpecificityThis 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 applicationsSuitable 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.

Properties

Applications

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.

Target

Anti-IRS2 (phospho S731) antibody images

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

    Secondary
    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

References for Anti-IRS2 (phospho S731) antibody (ab3690)

This product has been referenced in:
  • Chen J  et al. ZiBuPiYin recipe protects db/db mice from diabetes-associated cognitive decline through improving multiple pathological changes. PLoS One 9:e91680 (2014). WB ; Mouse . Read more (PubMed: 24614172) »
  • Lopez R  et al. Hyperglycemia enhances the proliferation of non-tumorigenic and malignant mammary epithelial cells through increased leptin/IGF1R signaling and activation of AKT/mTOR. PLoS One 8:e79708 (2013). WB ; Human . Read more (PubMed: 24260287) »

See all 4 Publications for this product

Product Wall

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

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 ...

Read More

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 ...

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"