701746

antibody from Invitrogen Antibodies

Targeting: IRS1 HIRS-1

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

701746

Provider

Invitrogen Antibodies

Product name

Phospho-IRS1 (Ser312) Recombinant Rabbit Monoclonal Antibody (18H26L6)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Monkey, Pig and Cat.

Antibody clone number

18H26L6

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) MCF7 (1), MCF7 treated with Insulin (100 nM Insulin for 10 min) (2), MCF7 treated with TNF alpha (50 ng TNF alpha for 20 min) (3), MCF7 treated with IGF-1 (100 ng IGF-1 for 20 min) (4), Hep G2 (5), Hep G2 treated with Insulin (100 nM Insulin for 10 min) (6). The blots were probed with Anti- IRS1 (pS312) Recombinant Rabbit Monoclonal Antibody (Product # 701746, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 180 kDa band corresponding to IRS1 (pS312) was observed. To confirm the specificity of Anti-IRS (pS312) Recombinant Rabbit Monoclonal Antibody, competition was performed with the phosphopeptide (10 µg/mL) as shown in the corresponding blot on the right. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with overnight wet transfer system. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP200

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) MCF7 (1), MCF7 treated with Insulin (100 nM Insulin for 10 min) (2), MCF7 treated with TNF alpha (50 ng TNF alpha for 20 min) (3), MCF7 treated with IGF-1 (100 ng IGF-1 for 20 min) (4), Hep G2 (5), Hep G2 treated with Insulin (100 nM Insulin for 10 min) (6). The blots were probed with Anti- IRS1 (pS312) Recombinant Rabbit Monoclonal Antibody (Product # 701746, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 180 kDa band corresponding to IRS1 (pS312) was observed. To confirm the specificity of Anti-IRS (pS312) Recombinant Rabbit Monoclonal Antibody, competition was performed with the phosphopeptide (10 µg/mL) as shown in the corresponding blot on the right. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with overnight wet transfer system. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP200