BODIPY FL Polyclonal Antibody, Invitrogen™

Description

• We offer a variety of anti-fluorescent dye antibodies that recognize specific fluorophores and, in most cases, quench their fluorescence
• These anti-dye antibodies, including our antibody to the BODIPYTM FL fluorophore, can serve as cell-impermeant probes for determining whether fluorescent dye-conjugated ligands, proteins, bacteria, or other biomolecules have been internalized by endocytic or pinocytic processes
• The anti-BODIPYTM FL antibody (A-5770) was raised against the BODIPY FL fluorophore and exhibits no appreciable cross-reaction with non-BODIPY dyes, although it will cross-react to varying extents with other BODIPY dyes
• In solution assays, we have found that this antibody effectively quenches the fluorescence of the BODIPY FL fluorophore, but quenches the BODIPY TR dye to a lesser extent and does not significantly quench the BODIPYTM TMR dye
• Invitrogen uses a sensitive quenching assay to ensure that this antibody is provided at a consistently high titer value
• As supplied, 20 μL of the antibody solution is certified to produce ≥50% of the maximal fluorescence quenching of 1 mL of a 50 nM solution of BODIPY FL dye, assayed in 100 mM sodium phosphate, pH 8.0
• Maximal quenching for BODIPY FL dye is ∽80% of the fluorescence of the free dye
• Due to steric hindrance, maximal fluorescence quenching of the BODIPY FL fluorophore covalently bound to protein may be significantly less
• We offer a variety of anti-fluorescent dye antibodies that recognize specific fluorophores and, in most cases, quench their fluorescence
• These anti-dye antibodies, including our antibody to the BODIPYTM FL fluorophore, can serve as cell-impermeant probes for determining whether fluorescent dye-conjugated ligands, proteins, bacteria, or other biomolecules have been internalized by endocytic or pinocytic processes
• The anti-BODIPYTM FL antibody (A-5770) was raised against the BODIPY FL fluorophore and exhibits no appreciable cross-reaction with non-BODIPY dyes, although it will cross-react to varying extents with other BODIPY dyes
• In solution assays, we have found that this antibody effectively quenches the fluorescence of the BODIPY FL fluorophore, but quenches the BODIPY TR dye to a lesser extent and does not significantly quench the BODIPYTM TMR dye
• Invitrogen uses a sensitive quenching assay to ensure that this antibody is provided at a consistently high titer value
• As supplied, 20 μL of the antibody solution is certified to produce ≥50% of the maximal fluorescence quenching of 1 mL of a 50 nM solution of BODIPY FL dye, assayed in 100 mM sodium phosphate, pH 8.0
• Maximal quenching for BODIPY FL dye is ∽80% of the fluorescence of the free dye
• Due to steric hindrance, maximal fluorescence quenching of the BODIPY FL fluorophore covalently bound to protein may be significantly less
• It is a good practice to centrifuge protein conjugate solutions briefly in a microcentrifuge before use; only the supernatant should then be added to the experiment
• This step will eliminate any protein aggregates that may have formed during storage, thereby reducing nonspecific background staining.