Western Blot (Cell lysate)
Western blot using IRS1 (phospho S307) polyclonal antibody (Cat # PAB10078) shows detection of a band at ~180 KDa believed to represent phosphorylated IRS1 (arrowhead).
Lane 1 shows staining of human 293 cell lysate.
Lane 2 shows staining of 293 cell lysate prepared from cells serum-starved for 18 h followed by treatment with 5 ug/mL of anisomysin for 30 min.
The pronounced staining of the band at 180 KDa is not seen when the antibody was pre-incubated with immunizing peptide prior to reaction (data not shown).
The identity of the intensely reactive bands at ~70 kD in both lane 1 and 2 is unknown, although these bands were also competedout by pre-incubation with the immunizing peptide.
Approximately 25 ug of each lysate was separated on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred onto nitrocellulose.
After blocking with 5% goat serum, 0.5%BLOTTO in PBS, the membrane was probed with the primary antibody diluted to 1:250.
Reaction occurred overnight at 4°C followed by washes and reaction with a 1:10,000 dilution of IRDye™800 conjugated Gt-a-RabbitIgG [H&L] MX for 45 min at room temperature (800 nmchannel, green).
Molecular weight estimation was made by comparison to prestained MW markers in lane M (700 nm channel, red).
IRDye™800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR.
IRDye is a trademark of LI-COR, Inc.
- Specifications
Product Description
Rabbit polyclonal antibody raised against synthetic phosphopeptide of IRS1.
Immunogen
Synthetic phosphopeptide corresponding to residues surrounding S307 of human IRS1.
Host
Rabbit
Reactivity
Bovine, Chicken, Dog, Human, Monkey, Mouse, Pig, Rat
Specificity
Reactivity occurs against human IRS1 pS307 protein and This antibody is specific to the phosphorylated form of the protein. Reactivity with non-phosphorylated human IRS1 is minimal by ELISA.
Form
Liquid
Quality Control Testing
Antibody Reactive Against Synthetic Peptide.
Recommend Usage
ELISA (1:2000-1:10000)
Western Blot (1:250-1:1500)
The optimal working dilution should be determined by the end user.Storage Buffer
In 20 mM KH2PO4, 150 mM NaCl, pH 7.2 (0.01% sodium azide)
Storage Instruction
Store at 4°C. For long term storage store at -20°C.
Aliquot to avoid repeated freezing and thawing.Note
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- Applications
Western Blot (Cell lysate)
Western blot using IRS1 (phospho S307) polyclonal antibody (Cat # PAB10078) shows detection of a band at ~180 KDa believed to represent phosphorylated IRS1 (arrowhead).
Lane 1 shows staining of human 293 cell lysate.
Lane 2 shows staining of 293 cell lysate prepared from cells serum-starved for 18 h followed by treatment with 5 ug/mL of anisomysin for 30 min.
The pronounced staining of the band at 180 KDa is not seen when the antibody was pre-incubated with immunizing peptide prior to reaction (data not shown).
The identity of the intensely reactive bands at ~70 kD in both lane 1 and 2 is unknown, although these bands were also competedout by pre-incubation with the immunizing peptide.
Approximately 25 ug of each lysate was separated on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred onto nitrocellulose.
After blocking with 5% goat serum, 0.5%BLOTTO in PBS, the membrane was probed with the primary antibody diluted to 1:250.
Reaction occurred overnight at 4°C followed by washes and reaction with a 1:10,000 dilution of IRDye™800 conjugated Gt-a-RabbitIgG [H&L] MX for 45 min at room temperature (800 nmchannel, green).
Molecular weight estimation was made by comparison to prestained MW markers in lane M (700 nm channel, red).
IRDye™800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR.
IRDye is a trademark of LI-COR, Inc.Immunohistochemistry
Enzyme-linked Immunoabsorbent Assay
- Gene Info — IRS1
Entrez GeneID
3667Protein Accession#
P35568;NP_005535Gene Name
IRS1
Gene Alias
HIRS-1
Gene Description
insulin receptor substrate 1
Gene Ontology
HyperlinkOther Designations
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- Interactomes
- Pathways
- Diseases
- Publication Reference
- Attenuation of insulin-stimulated insulin receptor substrate-1 serine 307 phosphorylation in insulin resistance of type 2 diabetes.
Danielsson A, Ost A, Nystrom FH, Stralfors P.
The Journal of Biological Chemistry 2005 Oct; 280(14):34389.
Application：WB-Ti, Human, Adipose tissue.
- Activation of the hexosamine pathway leads to phosphorylation of insulin receptor substrate-1 on Ser307 and Ser612 and impairs the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin insulin biosynthetic pathway in RIN pancreatic beta-cells.
Andreozzi F, D'Alessandris C, Federici M, Laratta E, Del Guerra S, Del Prato S, Marchetti P, Lauro R, Perticone F, Sesti G.
Endocrinology 2004 Mar; 145(6):2845.
- Salicylic acid reverses phorbol 12-myristate-13-acetate (PMA)- and tumor necrosis factor alpha (TNFalpha)-induced insulin receptor substrate 1 (IRS1) serine 307 phosphorylation and insulin resistance in human embryonic kidney 293 (HEK293) cells.
Jiang G, Dallas-Yang Q, Liu F, Moller DE, Zhang BB.
The Journal of Biological Chemistry 2003 Jan; 278(1):180.
Application：WB-Tr, Human, HEK 293 cells.
- Attenuation of insulin-stimulated insulin receptor substrate-1 serine 307 phosphorylation in insulin resistance of type 2 diabetes.