Our scientists are at the bench daily to produce and validate our antibodies, so they have hands-on experience and knowledge of each antibody’s performance.
In-Cell Western™ is a simple and cost effective means for quantification of intracellular signaling in whole cells. This assay involves seeding cells in microtiter plates followed by fixation/permeabilization and subsequent labeling with activation state-specific or control antibodies, infrared-conjugated secondary antibodies, and/or far-red DNA dye. The well-level data acquired by the LI-COR® Odyssey® Infrared Imaging System permit accurate measurement of signal induction or inhibition, with the potential for multiple treatments, end points, and replicates on each plate.
Antibodies for In-Cell Western™ Assays
Cell Signaling Technology (CST) offers over 1000 antibodies that may be used for In-Cell Western™ (ICW) assays. Because ICW assays are similar to classic immunocytochemistry, our antibodies that have been validated and approved for immunofluorescence (IF) are compatible with this assay. Simply follow our In-Cell Western™ Protocol to ensure correct labeling of your target proteins. If you have any questions, please contact our Technical Support team.
DyLight® Conjugated Secondary Antibodies
We offer secondary antibodies conjugated to DyLight® 680 or 800 near infrared fluorescent dyes. Due to their low background fluorescence, high sensitivity, photostability, and ease of quantification, DyLight® dyes are ideal for fluorescent western blotting and In-Cell Western™ (ICW) assays. Each CST DyLight® conjugated secondary antibody is tested in-house by fluorescent western and ICW analysis.
We also offer DRAQ5®, a near-infrared DNA dye. This dye can be used to normalize antibody signal to cell number, eliminating the potential for binding interference that may occur when multiplexing state-specific and total antibodies directed toward the same protein that have not been extensively tested for compatible use.