Applications

IRDye 800CW secondary antibodies can be used for numerous applications, including: Western blotting, In‑Cell Western™ Assay, In-Gel Western, and many others.

Two-color detection can be achieved by multiplexing IRDye 800CW secondary antibodies with IRDye 680RD or IRDye 680LT secondary antibodies. IRDye 800CW secondary antibodies are also suitable for immunofluorescent microscopy and other fluorescent imaging applications when using instrumentation with appropriate excitation and detection capabilities. Visit licor.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 800CW (MW 1166)

• Size: 0.1 mg

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

• Form of Antibody: IRDye 800CW purified immunoglobulin conjugate, lyophilized in phosphate buffered saline, pH 7.4. Contains 10 mg/mL BSA (IgG and protease free) as stabilizer and 0.01% sodium azide as preservative, after reconstitution.

  Contains sodium azide.

• Excitation Wavelength: 778 nm (in PBS)

• Emission Wavelength: 795 nm (in PBS)

• Immunogen: Rabbit IgG

Purity and Specificity

Isolation of specific antibodies was accomplished by affinity chromatography using pooled IgG covalently linked to agarose. Based on ELISA and flow cytometry, this antibody reacts with the heavy and light chains of rabbit IgG, and with the light chains of rabbit IgM and IgA. This antibody was tested by Dot Blot and/or solid-phase adsorbed for minimal cross-reactivity with human, mouse, rat, sheep, and chicken serum proteins, but may cross-react with immuno-globulins 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