IRDye® Secondary Antibodies Answer Portal
For example, "reconstitute secondary antibody".
Safety Data Sheets (SDS) are available on the SDS Search Page.
IRDye Secondary Antibodies are highly cross-adsorbed near-infrared fluorescent secondary antibodies. These antibodies are optimized for use with Odyssey Imagers and may be compatible with similar instruments.
Form: Lyophilized in phosphate-buffered saline, pH 7.4
Storage: Protect from light. Store at 4 °C before reconstitution.
Which antibody should I use?
IRDye 800CW Secondary Antibodies
Choose IRDye 800CW Secondary Antibodies for low background and high sensitivity in the 800 nm channel. Use these antibodies when detecting targets that require the highest sensitivity.
Recommended applications
Western Blot
In‑Cell Western™ Assay
On-Cell Western Assay
Protein Array
Immunohistochemistry
Small Animal Imaging
Microscopy
Tissue Section Imaging
Virus Titration Assay
IRDye 680RD Secondary Antibodies
Choose IRDye 680RD Secondary Antibodies for the lowest background and easiest optimization when detecting in the 700 nm channel.
Recommended applications
Western Blot
In‑Cell Western™ Assay
On-Cell Western Assay
Protein Array
Immunohistochemistry
Small Animal Imaging
Microscopy
Tissue Section Imaging
IRDye 680LT Secondary Antibodies
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.
Note: Please see the IRDye 680LT Antibody Pack Insert for specific detergents and protocol modifications needed for optimization.
Recommended applications
Western Blot
Immunohistochemistry
Microscopy
Tissue Section Imaging
Not recommended for Small Animal Imaging, In‑Cell Western Assay, or On-Cell Western Assay
Frequently Asked Questions
Why am I getting weak or no signal?
This could be due to several variables. It may be as simple as needing to purchase fresh antibody, but further troubleshooting with our support team may be required. The Western Blot Troubleshooting Form can help you determine the potential source.
The more details you provide, the better we can help you. So, fill out the form as completely as possible.
Can I freeze the secondary antibody after reconstitution?
This is not recommended, as freezing does not significantly extend the life of the secondary antibody.
Why does my image have black bands on a red or green background?
There are a couple of potential causes related to the wet lab protocol. It may be that too much protein was loaded or too much secondary antibody was used.
Do you have an anti-sheep secondary?
Donkey anti-goat cross-reacts strongly with sheep primary antibodies and can be used in place of an anti-sheep secondary antibody.
My secondary came as a powder; how do I make it liquid?
The secondary antibodies must be reconstituted before use.
Combine each 0.1 mg of secondary antibody with 0.1 mL of sterile distilled water.
Gently mix by inverting.
Allow the solution to stand at room temperature for at least 30 minutes before use. Protect the solution from light.
If the solution is not completely dissolved after standing at room temperature, centrifuge the solution.
When stored as recommended, this product is stable for 3 months.