Alkaline phosphatase activity in cell-free haemolymph samples was measured according to the method of Guardiola et al. (2014 (link)) with slight modifications. Aliquots of 100 µL of cell-free haemolymph were mixed with the same volume of 4 mM p-nitrophenyl liquid phosphate (Sigma) in 100 mM ammonium bicarbonate buffer containing 1 mM MgCl2 (pH 7.8, 30 °C). The OD was continuously measured at 405 nm at 1-min intervals over the course of 1 h in a plate reader (FLUOstar OPTIMA). One unit of activity was defined as the amount of enzyme required to release 1 µmol of p-nitrophenol phosphate product in 1 min and the activity was expressed as U mg−1 cell-free haemolymph proteins.
P nitrophenyl liquid phosphate
Manufactured by Merck Group
P-nitrophenyl liquid phosphate is a chemical compound used as a substrate in various enzymatic assays. It is a colorless to pale yellow liquid that undergoes a color change upon enzymatic hydrolysis, allowing for the detection and quantification of enzyme activity.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using p nitrophenyl liquid phosphate
1
Alkaline Phosphatase Activity in Hemolymph
2
Quantification of Alkaline Phosphatase
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Alkaline phosphatase activity was quantified as described by Mansour et al. (Mansour et al. 2017) . Aliquots of 100 µL of haemolymph were mixed with 100 µL of 4 mM p-nitrophenyl liquid phosphate (Sigma) in 100 mM ammonium bicarbonate buffer containing 1 mM MgCl2 (pH 7.8, 30 ºC). ALP activity was determined by recording the increase of OD at 405 nm every minute for 1 h. ALP activity was expressed as unit mg -1 haemolymph proteins, where one unit represents the amount of enzyme required to release 1 µmol of p-nitrophenol phosphate product in 1 min.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!