Applications

IRDye 680RD secondary antibodies can be used for numerous applications, including: Western blotting and the In‑Cell Western™ Assay. IRDye 680RD secondary antibodies are also suitable for immunofluorescent microscopy and other fluorescent imaging applications when using instrumentation with appropriate excitation and detection capabilities. Two-color detection can be achieved by multiplexing IRDye 680RD secondary antibodies with IRDye 800CW secondary antibodies. These antibodies are optimized for use with Odyssey family imagers.

Visit licorbio.com/support for protocols, example experiments, recommendations, and more technical information.

Storage

• Storage temperature: 4 °C
• Storage conditions: Protect from light and moisture.

• Shelf life: When stored as recommended, this product is stable for 3 months.

Specifications

• Fluorophore: IRDye 680RD (MW 1003)

• Size: 0.1 mg

• Antibody Concentration: 1.0 mg/mL (when reconstituted as directed)

• Form of Antibody: IRDye 680RD purified immunoglobulin conjugate, lyophilized in phosphate buffered saline, pH 7.4. Contains 10 mg/mL BSA (free of IgG and protease) as stabilizer and 0.01% sodium azide as preservative, after reconstitution.

  Contains sodium azide.

• Excitation Wavelength: 676 nm (in PBS)

• Emission Wavelength: 694 nm (in PBS)

• Immunogen: Goat IgG

The BSA used as a stabilizer for this product is free of IgG. However, blockers and antibody diluents made with bovine serum albumin (BSA) and dry milk may contain IgG that reacts with anti-bovine IgG, anti-goat IgG, anti-horse IgG, and anti-sheep IgG antibodies. This can lead to a significant increase in background and/or reduction of secondary antibody titer for protein detection applications.

Purity and Specificity

Isolation of specific antibodies was accomplished by affinity chromatography using pooled IgG covalently linked to agarose. Based on ELISA, this antibody reacts with the heavy and light chains of goat IgG and with sheep IgG. This antibody was tested by Dot Blot and/or solid-phase adsorbed for minimal cross-reactivity with human, mouse, rabbit, rat, chicken, guinea pig, hamster, swine, and horse serum proteins, but may cross-react with immunoglobulins from other species. The conjugate has been specifically tested and qualified for Western blot and In-Cell Western™ Assay applications.

Recommended Dilutions

Optimum dilutions will vary and should be determined empirically.

Avoid non-specific background staining on PVDF membranes: If you are making your own antibody diluent, add SDS (final concentration 0.01 – 0.02%) and Tween® 20 (final concentration 0.2%) to the diluent. If using Intercept® T20 Antibody Diluent, only add SDS (final concentration 0.01 – 0.02%).

Recommended Dilutions for Western blotting

Recommended Dilutions for the In‑Cell Western™ Assay