600-401-445

antibody from Rockland Immunochemicals, Inc.

Targeting: IRS1 HIRS-1

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

600-401-445

Provider

Rockland Immunochemicals, Inc.

Proper citation

Rockland Cat#600-401-445, RRID:AB_217958

Product name

Anti-IRS1 pS307 (RABBIT) Antibody - 600-401-445

Antibody type

Polyclonal

Vial size

100 µl

Western blot

Supportive validation

Submitted by

Rockland Immunochemicals, Inc. (provider)

Main image

Experimental details

Western blot using Rockland's Affinity Purified anti-IRS1 pS307 antibody 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 µg/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 competed out by pre-incubation with the immunizing peptide.  Approximately 25 µg 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-Rabbit IgG [H&L] MX (611-132-122) for 45 min at room temperature (800 nm channel, 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.  Other detection systems will yield similar results.

Validation comment

Western Blot