Invitrogen

IRS1 Monoclonal Antibody (K.346.8)

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

1 Reference

View all (74) IRS1 antibodies

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Datasheet

Protocols

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Tech Support

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IRS1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Insulin receptor substrate 1 was performed using 70% confluent log phase MIA PaCa-2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with IRS1 Monoclonal Antibody (K.346.8) (Product # MA5-15068) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained withRhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nucleus and cytoplasm localization. Panel e represents Panc-1. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60x magnification.

IRS1 Antibody in Western Blot (WB) — Western blot was performed using Anti-IRS1 Monoclonal Antibody (K.346.8) (Product # MA5-15068) and a ~130kDa band corresponding to Insulin receptor substrate 1 was observed across cell lines tested . Whole cell extracts (50 µg lysate) of MIA PaCa-2 (Lane 1), PANC-1 (Lane 2) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20000 using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).Relative expression observed between MIA Paca-2 and Panc-1.

IRS1 Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across MIA Paca-2 and Panc-1 owing to their inherent genetic constitution. Relative expression of Insulin receptor substrate 1 was observed ~130kDa in MIA Paca-2 using Anti-IRS1 Monoclonal Antibody (K.346.8) (Product # MA5-15068) in Western Blot. {RE}

Product Details

MA5-15068

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000	View 1 publication 1 publication
Immunocytochemistry (ICC/IF)	1:100	-

Product Specifications
Species Reactivity	Human, Mouse, Pig, Rat
Published species	Pig
Host/Isotype	Rabbit / IgG
Class	Monoclonal
Type	Antibody
Clone	K.346.8
Immunogen	Synthetic peptide corresponding to the sequence surrounding Ser307 of human IRS-1 View antigen
Conjugate	Unconjugated
Form	Liquid
Purification	Affinity chromatography
Storage buffer	0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/mL BSA, 50% glycerol
Contains	<0.02% sodium azide
Storage conditions	-20°C
RRID	AB_10988670

Product Specific Information

It is not recommended to aliquot this antibody.

This antibody is not cross-reactive with IRS-2.

Target Information

IRS-1, a major substrate of the insulin receptor, is phosphorylated in response to stimulation of cells by insulin, insulin-like growth factor 1 (IGF-1) and interleukin 4 (IL-4). IRS-1 is phosphorylated on serine, threonine and tyrosine residues in a variety of tissues. An insulin-sensitive serine/threonine kinase casein kinase II mediates a portion of the insulin-stimulated serine/threonine phosphorylation of overexpressed IRS-1 in vivo. Thr 502 is identified as the major casein kinase II-catalyzed phosphorylation site in rat IRS-1, and Ser 99 is an additional phosphorylation site catalyzed by casein kinase II. Thus, casein kinase II-catalyzed phosphorylation of IRS-1 may be a component of the intracellular insulin signaling cascade. IRS-1 contains three putative binding sites for 14-3-3 (Ser 270, Ser 374 and Ser 641) and the motif around Ser 270 is located in the phosphortyrosine binding domain of IRS-1, which is responsible for the interaction with the insulin receptor.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Berberine induces lipolysis in porcine adipocytes by activating the AMP‑activated protein kinase pathway.

Molecular medicine reports

Yang Y,Lu R,Gao F,Zhang J,Liu F

MA5-15068 was used in Western Blot to suggest that BBR induced lipolysis in porcine adipocytes via a pathway that involves AMPK activation, but does not cause insulin resistance.