Biotinylation

1. It is important to remember that biotinylation depends on initial concentration of protein. The level of biotinylation is increased at a higher concentration of protein even at same NHS-LC-biotin:protein molar ratio.

2. Biotinylation efficiency and its effects on protein activities vary significantly from protein to protein. In case of catalase, biotinylation does not change the catalytic activity at up to a level of 4-5 biotin/catalase monomer.

Catalase Cell Culture H2o2

Fig. 5. Cell protection against H2O2-induced oxidative stress by catalase/anti-PECAM immunoconjugates. Cells were treated with the immunoconjugates as described in Subheading 3.4.2. at several concentrations. (A) Degradation of H2O2 by bound catalase-containing conjugates. Initial concentrations of immunoconjugates used for cell treatment are indicated in micrograms of catalase/well. (B) Cell death as a result of severe oxidative stress was analyzed by release of [51Cr]. Cells were incubated with 5 mM H2O2 for 5 h. Release of [51Cr] in control cells shows level of passive diffusion whereas practically complete [51Cr] release in the absence of immunoconjugates demonstrates significant cell death. (C) Phase-contrast microscopy of control and treated cells.

Fig. 5. Cell protection against H2O2-induced oxidative stress by catalase/anti-PECAM immunoconjugates. Cells were treated with the immunoconjugates as described in Subheading 3.4.2. at several concentrations. (A) Degradation of H2O2 by bound catalase-containing conjugates. Initial concentrations of immunoconjugates used for cell treatment are indicated in micrograms of catalase/well. (B) Cell death as a result of severe oxidative stress was analyzed by release of [51Cr]. Cells were incubated with 5 mM H2O2 for 5 h. Release of [51Cr] in control cells shows level of passive diffusion whereas practically complete [51Cr] release in the absence of immunoconjugates demonstrates significant cell death. (C) Phase-contrast microscopy of control and treated cells.

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  • leah
    How much level of h2o2 in cells?
    8 years ago
  • aman gabriel
    Does biotinylation effect protein folding?
    8 years ago

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