IRS1 antibody (N-Term)

Catalog No. ABIN3042564

Product Details anti-IRS1 Antibody

Target: IRS1 (Insulin Receptor Substrate 1 (IRS1))

Binding Specificity: AA 106-122, N-Term

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This IRS1 antibody is un-conjugated

Application: Western Blotting (WB)

Purpose: Rabbit IgG polyclonal antibody for Insulin receptor substrate 1(IRS1) detection. Tested with WB in Human,Mouse,Rat.

Sequence: WYQALLQLHN RAKAHHD

Cross-Reactivity (Details): Predicted Cross Reactivity: mouse, rat
No cross reactivity with other proteins.
Predicted Cross Reactivity: Species predicted to be fit for the product based on sequence similarities.

Characteristics: Rabbit IgG polyclonal antibody for Insulin receptor substrate 1(IRS1) detection. Tested with WB in Human,Mouse,Rat.
Gene Name: insulin receptor substrate 1
Protein Name: Insulin receptor substrate 1

Purification: Immunogen affinity purified.

Immunogen: A synthetic peptide corresponding to a sequence at the N-terminus of mouse IRS1(106-122aa WYQALLQLHNRAKAHHD), identical to the related rat sequence, and different from the related human sequence by one amino acid.

Isotype: IgG

Application Details

Application Notes: WB: Concentration: 0.1-0.5 μg/mL, Tested Species: Human, Predicted Species: Mouse, Rat
Notes: Tested Species: Species with positive results. Predicted Species: Species predicted to be fit for the product based on sequence similarities.
Other applications have not been tested. Optimal dilutions should be determined by end users.

Comment:

Antibody can be supported by chemiluminescence kit ABIN921124 in WB.

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #300033 (Immunohistochemistry)

by: Human Protein Atlas

No.: #300033

Date: 12/13/2016

Antigen: IRS1

Method validated: Immunohistochemistry

Positive Control: colon, tonsil

Notes: Passed. ABIN3042564 staining is consistent with RNA-seq data and published literature.

Validation Images

Staining of human colon (A.) and tonsil (B.) sections in a tissue microarray with ABIN3042564. See the protocol for detailed information.

Full Methods

Primary Antibody: ABIN3042564

Secondary Antibody: HRP polymer (ThermoFisher Scientific, TL-125-PH)

Full Protocol:

• Sample preparation:
• • Fixation and TMA preparation according to PMID: 22688270.
  • Cut 4µm TMA sections using a waterfall microtome (ThermoFisher Scientific, HM355S).
  • Dry sections ON at RT and then back them for 12-24h at 50°C.
• Deparaffinization and rehydration in xylene and graded alcohol series:
• • xylene twice for 3min.
  • 100% ethanol for 1min.
  • 95% ethanol for 1min.
  • 0.3% H₂O₂ in 95% ethanol for 5min to block endogenous peroxidase.
  • 70% ethanol for 1min.
  • 50% ethanol for 1min.
  • distilled H₂O for 2min.
• Antigen Retrieval by Heat Induced Epitope Retrieval (HIER):
• • Boil the sections in Lab Vision Citrate buffer pH6.0 (ThermoFisher Scientific, TA-250-PM1X) for 4min at 125°C in a decloaking chamber (Biocare Medical).
  • Allow slides to cool down to 90°C in the decloaking chamber.
• Immunostaining in a Lab Vision Autostainer 480 (ThermorFisher Scientific) at RT with 300µl of reagents for each step:
• • Rinse sections in Lab Vision wash buffer with extra tween added to a final concentration of 0.2% (ThermoFisher Scientific, TA-999-TT and TA-125-TW) wash buffer.
  • Block sections with Lab Vision Ultra V-Block (ThermoFisher Scientific, TA-125-UB) for 5min.
  • Rinse sections 2x in wash buffer.
  • Incubate sections with primary rabbit anti-IRS1 antibody (antibodies-online, ABIN3042564) diluted 1:1100 for 30min.
  • Rinse sections 3x in wash buffer.
  • Incubate sections with labeled HRP polymer (ThermoFisher Scientific, TL-125-PH) for 30min.
  • Rinse sections 2x in wash buffer.
  • Develop in DAB Quanto solution (ThermoFisher Scientific, TL-125-QHDX) for 5min.
  • Rinse sections in distilled H₂O.
• Washing, counterstaining, and coverslipping in Autostainer XL (Leica Biosystems, ST5010) at RT:
• • Counterstaining in Mayer’s hematoxylin plus (HistoLab, 01820) for 7.5min.
  • Rinse sections in tap water for 5min.
  • Rinse sections in lithium carbonate water diluted 1:5 for 5min.
  • Dehydration in graded ethanol series and Neo-Clear (Merck Millipore, 109843) according to the manufacturer recommendations.
  • Automated mounting (Leica Biosystems, CV5030) of coverslip with Pertex (Histolab, 00871).
• Microscopy:
• • Image acquisition on a Leica Aperio AT2 and Aperio Scanscope AT at 20x magnification.

Experimental Notes: ABIN3042564 shows nice IRS1 staining in endothelial cells in most tissues as is supported by both literature and RNA-seq data. At lower dilutions the antibody does also show some cytoplasmic staining in the tested tissues.

Handling

Format: Lyophilized

Reconstitution: Add 0.2 mL of distilled water will yield a concentration of 500 μg/mL.

Concentration: 500 μg/mL

Buffer: Each vial contains 5 mg BSA, 0.9 mg NaCl, 0.2 mg Na2HPO4, 0.05 mg Thimerosal, 0.05 mg Sodium azide.

Preservative: Thimerosal (Merthiolate), Sodium azide

Precaution of Use: This product contains Sodium azide and Thimerosal (Merthiolate): POISONOUS AND HAZARDOUS SUBSTANCES which should be handled by trained staff only.

Handling Advice: Avoid repeated freezing and thawing.

Storage: 4 °C/-20 °C

Storage Comment: At -20°C for one year. After reconstitution, at 4°C for one month.
It can also be aliquotted and stored frozen at -20 °C for a longer time. Avoid repeated freezing and thawing.

References for anti-IRS1 antibody (ABIN3042564)

Mi, Chen, Zheng, Li, Zhang, Mo, Yang: "MicroRNA-139-5p Suppresses 3T3-L1 Preadipocyte Differentiation Through Notch and IRS1/PI3K/Akt Insulin Signaling Pathways." in: Journal of cellular biochemistry, Vol. 116, Issue 7, pp. 1195-204, (2015) (PubMed).

Target Details for IRS1

Target: IRS1 (Insulin Receptor Substrate 1 (IRS1))

Alternative Name: IRS1 (IRS1 Products)

Synonyms: HIRS-1 antibody, G972R antibody, IRS-1 antibody, IRS1IRM antibody, irs1 antibody, irsu antibody, IRS1 antibody, irs-1 antibody, irs1-a antibody, insulin receptor substrate 1 antibody, insulin receptor substrate 2 antibody, insulin receptor substrate 1 S homeolog antibody, IRS1 antibody, Irs1 antibody, irs2 antibody, irs1.S antibody

Background: Insulin receptor substrate 1(IRS-1) is a signalling adapter protein that in humans is encoded by the IRS-1 gene. It is mapped to 2q36.3. This gene exhibited no intrinsic enzyme activity, and it can serve as a docking protein involved in binding and activating other signal transduction molecules after being phosphorylated on tyrosine by insulin receptor kinase. IRS1 plays a key role in transmitting signals from the insulin and insulin-like growth factor-1(IGF-1) receptors to intracellular pathways PI3K/Akt and Erk MAP kinase pathways. IRS1 also plays important biological function for both metabolic and mitogenic(growth promoting) pathways. In addition to those, IRS1 is a key regulator of PI3K within malignant cells.

Synonyms: HIRS 1 antibody|HIRS1 antibody|Insulin Receptor Substrate 1 antibody||IRS 1 antibody|IRS-1 antibody|IRS1 antibody|OTTHUMP00000164234 antibody

UniProt: P35569

Pathways: Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Positive Regulation of Peptide Hormone Secretion, Hormone Transport, Negative Regulation of Hormone Secretion, Response to Growth Hormone Stimulus, Carbohydrate Homeostasis, Regulation of Carbohydrate Metabolic Process