What substrate does alkaline phosphatase use?
Substrates for Alkaline Phosphatase Nitroblue Tetrazolium (NBT) is used with the alkaline phosphatase substrate 5-Bromo- 4-Chloro-3-Indolyl Phosphate (BCIP) in western blotting and immunohistological staining procedures.
Is BCIP light sensitive?
The substrate is light sensitive and must be protected from light during incubation with the alkaline phosphatase-labeled probe.
Which reaction is catalyzed by the enzyme alkaline phosphatase?
Alkaline phosphatase catalyzes the hydrolysis of glucose-6-phosphate via a dissociative mechanism.
What is the cellular reaction that the enzyme alkaline phosphatase catalyzes?
Alkaline phosphatase encompasses a family of enzymes that catalyze the hydrolysis of organic phosphate esters, optimally at alkaline pH. Cell membranes of many cell types have ALP activity; however, only a few produce sufficient ALP to affect serum activity.
What type of enzyme is alkaline phosphatase?
homodimeric protein enzyme
Alkaline phosphatase (ALP, ALKP, ALPase, Alk Phos) (EC 3.1. 3.1), or basic phosphatase, is a homodimeric protein enzyme of 86 kilodaltons. Each monomer contains five cysteine residues, two zinc atoms and one magnesium atom crucial to its catalytic function, and it is optimally active at alkaline pH environments.
At what pH does alkaline phosphatase denature?
Both acid and alkaline phosphatases exist in living tissues. Alkaline phosphatase works best when the pH is about 8.6. An acid pH (below 7.0) hampers the activity of alkaline phosphatase, and a pH below 5.0 might denature it, according to the Journal of Bacteriology.
How do you prevent protein degradation in western blot?
How To Preserve Your Samples In Western Blotting
- Work quickly. Working quickly can minimize potential damage to your samples by simply allowing less time for them to become degraded.
- Keep everything cool. Heat is the enemy of proteins in solution, since proteases are active at warmer temperatures.
- Use protease inhibitors.