IRDye^® Secondary Antibodies

Highly Cross-Adsorbed IRDye Secondary Antibodies Provide Superior Performance

Quality and performance of secondary antibodies is crucial for Western blotting, In-Cell Western™ Assays, immunohistochemistry, and many other applications. IRDye Secondary Antibodies are highly cross-adsorbed, making them suitable for multi-color detection.

IRDye Secondary Antibodies have been optimized for use with the Odyssey^® family of imaging systems and can be used on instruments with similar excitation and emission filters.

Choosing the Right Dye-Conjugated Antibody

Select the right IRDye Secondary Antibody for your application using information in the table below. Here are some general guidelines to help you choose the right secondary antibodies:

• Choose IRDye 800CW Secondary Antibodies for low background and high sensitivity in the 800 nm channel. Use IRDye 800CW for detecting targets that require the highest sensitivity. Dilution working range is 1:5,000 – 1:25,000 for Western blots and 1:200 – 1:1,200 for In-Cell Western Assays. Other assays will require user optimization.
• Choose IRDye 680RD Secondary Antibodies for the lowest background and easiest optimization when detecting in the 700 nm channel. Dilution working range is 1:5,000 – 1:25,000 for Western blots and 1:200 – 1:1,200 for In-Cell Western Assays. Other assays will require user optimization.
• Choose IRDye 680LT Secondary Antibodies for applications that require greater light stability (such as microscopy) and when optimal Western blot conditions for specific targets have already been well-established. IRDye 680LT Secondary Antibodies tend to require more optimization than IRDye 680RD Secondary Antibodies, but they have the advantage of typically being used at much lower concentrations. Dilution range is 1:10,000 – 1:50,000 for Western blots. Other assays will require user optimization. Note: Please see the IRDye 680LT antibody pack insert for specific detergents and protocol modifications needed for optimization.