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 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 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: Goat IgG, whole molecule

Purity and Specificity

The antibody was isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. Based on immunoelectrophoresis, the antibody reacts with the heavy chains on goat IgG and with the light chains common to most goat immunoglobulins. The antibody also reacts with sheep IgG. No antibody was detected against non-immuno-globulin serum proteins. This antibody has been tested by ELISA and/or solid-phase adsorbed to ensure minimal cross-reaction with chicken, guinea pig, Syrian hamster, horse, human, mouse, rabbit, and rat serum proteins, but the antibody may cross-react with immunoglobulins from other species. The conjugate has been specifically tested and qualified for Western blot 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