Product Details
| Cat # +Size | K353-100 |
|---|---|
| Size | 100 assays |
| Kit Summary | BioVision’s Maltose Phosphorylase Activity Assay Kit is the first commercial assay to determine the Maltose Phosphorylase activity in different bacterial lysates. The assay converts maltose to produce Glucose, which is then detected by a set of enzymatic reactions to generate a fluorescent product with a Ex/Em 535/587 nm. The fluorescence signal is directly proportional to the Maltose Phosphorylase activity. BioVision’s Matlose Phosphorylase Activity Assay Kit is rapid, sensitive and a convenient tool for detecting Maltose Phosphorylase activity. It can detect as low as 0.5 mU under the assay conditions. |
| Detection Method | Fluorescence (Ex/Em 535/587 nm) |
| Species Reactivity | Bacterial lysates |
| Applications | • Analysis of Maltose Phosphorylase kinetics in inhibition or activation screening • Measurement of Maltose Phosphorylase activity in different bacteria |
| Features & Benefits | • Sensitive • Rapid • Convenient tool for detecting Maltose Phosphorylase activity • It can detect as low as 0.5 mU under the assay conditions |
| Kit Components | • Maltose Assay Buffer • Maltose Developer • Maltose • Maltose Probe • Glucose Standard • Maltose Phosphorylase |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Maltose phosphorylase (EC 2.4.1.8) belongs to the family of hexosyltransferases. It catalyzes maltose and inorganic phosphate to produce glucose and glucose-1-phosphate (G1P) during Maltose metabolism. Maltose phosphorylase is commonly identified in gram
positive and gram negative bacteria including E. coli, Bacillus subtilis, Enterococcus sp. and Lactobacillus brevis etc. BioVision’s Maltose Phosphorylase Activity Assay Kit is the first commercial assay to determine the Maltose Phosphorylase activity in different bacterial
lysates. The assay converts maltose to produce Glucose, which is then detected by a set of enzymatic reactions to generate a fluorescent product with a Ex/Em 535/587 nm. The fluorescence signal is directly proportional to the Maltose Phosphorylase activity. BioVision’s
Matlose Phosphorylase Activity Assay Kit is rapid, sensitive and a convenient tool for detecting Maltose Phosphorylase activity. It can detect as low as 0.5 mU under the assay conditions.
Maltose Phosphorylase Detection Solution Maltose + Pi Glucose Fluorescent Product (Ex/Em = 535/587 nm)
II. Application:
• Measurement of Maltose Phosphorylase activity in different bacteria
• Analysis of Maltose Phosphorylase kinetics in inhibition or activation screening.
Product name
Detection method
Sample type
Assay type
Assay duration
Product overview
Maltose Phosphorylase Assay Kit (Fluorometric) (ab273317) is the first commercial assay to determine the Maltose Phosphorylase activity in different bacterial lysates. The assay converts maltose to produce Glucose, which is then detected by a set of enzymatic reactions to generate a fluorescent product with a Ex/Em 535/587 nm. The fluorescence signal is directly proportional to the Maltose Phosphorylase activity.
This Kit is rapid, sensitive and a convenient tool for detecting Maltose Phosphorylase activity. It can detect as low as 0.5 mU under the assay conditions.
Notes
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Storage Conditions and Reagent Preparation:
Store the kit at -20°C. The kit components are stable for one year when stored as recommended. Briefly centrifuge small vials at low speed prior to opening. Read the entire protocol before performing the experiment.
Maltose Assay Buffer: Ready to use as supplied. Warm bottle to room temperature (RT) before use. Store at 4°C.
Maltose Probe (in DMSO): Ready to use as supplied. Warm to RT prior to use to melt frozen DMSO. Store at -20°C, protected from
light and moisture. Use within two months.
Maltose Developer and Maltose: Reconstitute each component with 220 µl of Assay Buffer. Store at -20°C. Keep on ice while in use.
Use within two months.
Glucose Standard: Store at -20°C. Warm to RT before use.
Maltose Phosphorylase: Add 450 μl of 50% glycerol (not included) to the vial to prepare the enzyme stock. Vortex to mix. Aliquot and
store at -20°C. Avoid multiple freeze-thaw cycles. Use within two months.
VII. Maltose Phosphorylase Activity Assay Protocol:
1. Sample Preparation: Grow cells in a suitable growth medium at 37°C overnight. After incubation, centrifuge at 10,000 x g for 20 min to harvest the cells and measure the weight of the pellet. Add 5 ml of ice-cold PBS into the cell pellet and disperse the pellet. Sonicate the cells for 5-10 min on ice. After sonication, centrifuge the cells at 10,000 x g for 30 min and transfer the supernatant to a new tube. For Sample and Background wells, add 2 μl of the cell supernatant to the desired well(s) in a 96-well black flat-bottom plate. Adjust the volume to 50 μl using Maltose Assay Buffer.
For Positive Control, prepare a 10 fold dilution of the Maltose Phosphorylase enzyme by adding 5 μl of the enzyme with 45 μl of Maltose Assay Buffer. Add 2 μl of the diluted enzyme into the Positive Control well. Adjust the volume to 50 μl using Maltose Assay Buffer.
2. Standard Curve Preparation: Mix 10 µl of Glucose Standard with 990 µl of Assay Buffer to prepare 1 mM diluted Glucose Standard. Mix 100 µl of 1 mM diluted Glucose Standard with 900 µl of Assay Buffer to prepare 0.1 mM diluted Glucose Standard solution. Add 0,4, 8, 12, 16 and 20 μl of the 0.1 mM diluted Standard into the desired wells to generate 0, 0.4, 0.8, 1.2, 1.6 and 2.0 nmole of Standard/well respectively. Adjust the volume to 50 μl using Maltose Assay Buffer.
