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.

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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: Rabbit IgG

Purity and Specificity

The antibody was isolated by affinity chromatography using antigens coupled to agarose beads. Based on ELISA, this antibody reacts with the heavy and light chains of rabbit IgG and with the light chains common to most rabbit immunoglobulins. This antibody has been tested by ELISA and/or solid-phase adsorbed to ensure minimal cross-reaction with bovine, chicken, goat, guinea pig, hamster, horse, human, mouse, rat, and sheep serum proteins, but the antibody 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