Slides were then incubated in ABC (avidin
biotin) reagent and washed in TBS buffer again for 2-5 minutes.
Biotin and Avidin
were purchased from Macklin Biochemical Co.
Cross-linker biotin binding proteins such as Avidin
, Streptavidin, and NeutrAvidin can be attached to the biotinylated PLGA NP.
The free avidin
and antibodies were removed by washing with PBS.
The iminobiotin coupled with anticancer drugs shows a high affinity to avidin
(and avidin-immobilized magnetic nanoparticles (AMNPs)) at elevated pH (9.5-10.8) but releases the drugs easily at lower pH (~4) or under the presence of a certain concentration of biotin at pH 6.8 according to the manufacturer's instructions.
When ABC-HRP was used, endogenous biotin reactivity was blocked by supplementing the 5% goat serum (Sigma) during blocking with 1: 20 avidin
and the primary antibody staining solution with 1 : 20 biotin (Avidin-biotin blocking kit; Vector Labs), with 3 x 2min washes with PBS in between.
Then, sections were blocked with 10% normal serum from the species of secondary antibody, Avidin
D solution (Avidin/Biotin Blocking Kit, Vector, Burlingame, CA, USA) and 0.25% Triton X-100 for 30 min.
(6) The biotin molecule, which binds irreversibly to avidin
, can be conjugated to an immunoglobulin molecule by a straightforward amino substitution at an alkaline pH with no loss of immunoglobulin activity.
EW also contain avidin
complexes with biotin to impair its digestion , which can lead to a biotin deficiency if consumed in large amounts that exceed the capacity of pepsin [20, 28].
The PCR solution and carbon particles conjugated with avidin
were directly added to the nitrocellulose membrane with two test lines of immobilized antibodies specific for digoxigenin and fluorescein.
(53) The size of avidin
with the molecular mass of 70 kDa is ~5 nm in diameter, which is large enough to be visualized directly by electron microscopy.
Comparison of avidin
biotin ELISA with RBPT and STAT for screening of antibodies to bovine brucellosis.