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Alkaline Phosphatase (ALP) Activity Assay Kit (PNPP Method)

Cat.No.:E-BC-K009-M
Detection instrument: Microplate reader(405 nm-415 nm,optimum wavelength: 405 nm)

Detection method: Colorimetric method

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500 Assays

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Detection principle

Alkaline phosphatase (ALP) is a group of cytomembrane-related enzymes with hydrolysis and transfer activity, acting on a variety of phosphate substrates. ALP is a homologous dimerase and each catalytic site contains three metal ions. There are four isozymes in humans: tissue nonspecific ALP, intestinal ALP, placental ALP and genital cell ALP.

Performance characteristics

Synonyms	ALP
Sample type	Serum, plasma, animal tissue, cells
Sensitivity	0.27 U/L
Detection range	0.27-50.8 U/L
Detection method	Colorimetric method
Assay type	Enzyme Activity
Assay time	30 min
Precision	Average inter-assay CV: 4.600%Average intra-assay CV: 1.200%
Other instruments required	Micropipettor, Water bath, Incubator
Other reagents required	Normal saline (0.9% NaCl)
Storage	-20℃
Valid period	12 months

Dilution of sample

It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (0.27-50.8 U/L).

The recommended dilution factor for different samples is as follows (for reference only):

Sample type	Dilution factor
Human serum	10-20
Mouse serum	20-30
Rat serum	10-20
Mouse plasma	10-20
HepG2 supernatant	8-12
10% Rat kidney tissue homogenate	400-1200
10% Mouse liver tissue homogenate	10-15
10% Rat lung tissue homogenate	50-100
10% Mouse brain tissue homogenate	30-50
Human urine	1

Note: The diluent is reagent 1.

Troubleshooting

Collection and treatment of common samples for metabolism assay detection

Questions and Answers of Metabolism Assay Kits

Citations

Science Advances (2019) IF: 14.143
A prokaryotic-eukaryotic hybrid viral vector for delivery of large cargos of genes and proteins into human cells

DOI: 10.1126/sciadv.aax0064

Sample: serum
ENVIRONMENTAL SCIENCE & TECHNOLOGY (2022) IF: 9.028
Lipid Metabolic Disorder Induced by Pyrethroids in Nonalcoholic Fatty Liver Disease of Xenopus laevis

DOI: 10.1021/acs.est.2c00516

Sample: plasma
JOURNAL OF HAZARDOUS MATERIALS (2020) IF: 9.038
The combined adverse effects of cis-bifenthrin and graphene oxide on lipid homeostasis in Xenopus laevis

DOI: 10.1016/j.jhazmat.2020.124876

PMID: 33360192
JOURNAL OF CELLULAR PHYSIOLOGY (2022) IF: 6.513
IGF2BP2, an RNA-binding protein regulates cell proliferation and osteogenic differentiation by stabilizing SRF mRNA

DOI: 10.1002/jcp.30919
Frontiers in Endocrinology (2021) IF: 5.555
Chondroitin Sulfate Alleviates Diabetic Osteoporosis and Repairs Bone Microstructure via Anti-Oxidation, Anti-Inflammation, and Regulating Bone Metabolism

DOI: 10.3389/fendo.2021.759843

PMID: 34777254
Sample: serum
Frontiers in Pharmacology (2021) IF: 5.811
Phillygenin Attenuates Carbon Tetrachloride-Induced Liver Fibrosis via Modulating Inflammation and Gut Microbiota

DOI: 10.3389/fphar.2021.756924

PMID: 34621179
Sample: serum
RARE METALS (2021) IF: 4.003
Degradation and biological performance of porous osteomimetic biphasic calcium phosphate in vitro and in vivo

DOI: 10.1007/s12598-021-01814-0

Sample: MG63 cell
BONE (2021) IF: 4.398
Coupling Induction of Osteogenesis and Type H Vessels by Pulsed Electromagnetic Fields in Ovariectomy-induced Osteoporosis in Mice

DOI: 10.1016/j.bone.2021.116211

PMID: 34560308
Sample: serum
Scientific Reports (2022) IF: 4.996
Endothelin receptor B enhances liver injury and pro-inflammatory responses by increasing G-protein-coupled receptor kinase-2 expression in primary biliary cholangitis

DOI: 10.1038/s41598-022-21816-x

Sample: serum
NEUROSCIENCE LETTERS (2021) IF: 3.046
Syringic acid protects against thioacetamide-induced hepatic encephalopathy: Behavioral, biochemical, and molecular evidence

DOI: 10.1016/j.neulet.2021.136385

PMID: 34871743
Sample: serum
BMC MUSCULOSKELETAL DISORDERS (2022) IF: 2.562
miR-215-5p regulates osteoporosis development and osteogenic differentiation by targeting XIAP

DOI: 10.1186/s12891-022-05731-w

PMID: 35978328
Sample: hBMSC
Evidence-based Complementary and Alternative Medicine (2022) IF: 2.65
San-Huang-Chai-Zhu Formula Ameliorates Liver Injury in Intrahepatic Cholestasis through Suppressing SIRT1/PGC-1α-Regulated Mitochondrial Oxidative Stress

DOI: 10.1155/2022/7832540

Sample: hepatic tissue,serum
Journal of Oleo Science (2022) IF: 1.628
Chemoprotective Effect of Decalactone on Hepatic Cancer via Diminishing the Inflammatory Response and Oxidative Stress

DOI: 10.5650/jos.ess22033

PMID: 35965085
Sample: liver
Toxicology Reports (2022)
Effect of silymarin on blood coagulation profile and osmotic fragility in carbon tetrachloride induced hepatotoxicity in male Wistar rats

DOI: 10.1016/j.toxrep.2022.06.005

Sample: plasma
Toxicology Reports (2017)
Diclofenac – induced hepatotoxicity: Low dose of omega-3 fatty acids have more protective effects

DOI: 10.1016/j.toxrep.2017.12.002

Sample: plasma

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Reviews/Q&A

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Q&A

V**************oSubmitted [ Oct 05 2023 ]

Asked: How total cellular proteins are measured?

adminSubmitted [ Oct 05 2023 ]

Answered: The following are the detailed steps of how the cell samples were processed and how the protein concentration was determined. ① Harvest the number of cells needed for each assay (initial recommendation =3 × 10^6 cells). ② Wash cells with 0.9% NaCl . ③ Homogenize 3 × 10^6 cells in 200-400 µL 0.9% NaCl (μL) with a ultrasonic cell disruptor at 4℃. ④ Centrifuge at 10000 × g for 10 minutes to remove insoluble material. Collect supernatant and keep it on ice for detection. ⑤ Meanwhile, determine the protein concentration of supernatant (E-BC-K318-M).

R************sSubmitted [ Nov 12 2021 ]

Asked: Dear Sir, Madame, I was wondering which are the units of this assay? Units per L in glycine buffer? Kind Regards, Raquel Martins

adminSubmitted [ Nov 12 2021 ]

Answered: 1. ALP activity in Serum (plasma) sample Definition: The amount of 1 μmol p-nitrophenol produced by 1 L serum (plasma) per minute catalyzing the substrate at 37℃ is defined as 1 activity unit. Calculation formula: ALP activity (U/L) = (ΔA-b) ÷ a ÷ T × f 2. ALP activity in the tissue sample Definition: The amount of 1 μmol p-nitrophenol produced by 1 g tissue protein per minute atalyzing the substrate at 37℃ is defined as 1 activity unit. Calculation formula: ALP activity (U/gprot) = (ΔA - b) ÷ a ÷ T × f ÷ Cpr 3. ALP activity in the cell sample Definition: The amount of 1 μmol p-nitrophenol produced by 1 g cell protein per minute catalyzing the substrate at 37℃ is defined as 1 activity unit. Calculation formula: ALP activity (U/gprot) = (∆A - b ) ÷ a ÷ T × f ÷ Cpr

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