Serum was thawed and analyzed for the Ca content using a microplate reader with Ca assay kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). Serum P contents were determined by the molybdenum blue method (Goldenberg and Fernandez, 1966 (link)). The 25-OHD3 level in serum was determined by the method of ELISA with the assay kits (Nanjing Jiancheng Bioengineering Institute). The left tibias were grinded, homogenized, and then sonicated and centrifuged at 1,000 × g for 10 min at 4°C to harvest the supernatants for ALP activity analysis. The ALP activities in serum and the tibia were measured using a microplate reader with ALP assay kits (Nanjing Jiancheng Bioengineering Institute).
Alp assay kit
Manufactured by Nanjing Jiancheng
Sourced in China
The ALP assay kit is a laboratory tool used to measure the activity of the enzyme alkaline phosphatase (ALP) in biological samples. ALP is an important enzyme involved in various physiological processes and its measurement can provide valuable information for research and clinical applications.
Automatically generated - may contain errors
Lab products found in correlation
Bca protein assay kit, by Thermo Fisher Scientific (8 mentions)
Bca protein assay kit, by Beyotime (8 mentions)
Nbt bcip staining kit, by Beyotime (8 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (8 mentions)
Bcip nbt alkaline phosphatase color development kit, by Beyotime (7 mentions)
Nbt bcip staining kit, by CoWin Biotech (6 mentions)
Dexamethasone (dex), by Merck Group (6 mentions)
Bcip nbt alp color development kit, by Beyotime (5 mentions)
Ripa lysis buffer, by Beyotime (4 mentions)
β glycerophosphate, by Merck Group (4 mentions)
Bca assay kit, by Beyotime (4 mentions)
Ascorbic acid, by Merck Group (4 mentions)
Pierce protein assay kit, by Thermo Fisher Scientific (3 mentions)
Alizarin red buffer, by Merck Group (3 mentions)
Ca assay kit, by Nanjing Jiancheng (2 mentions)
L ascorbic acid, by Merck Group (2 mentions)
Ripa buffer, by Beyotime (2 mentions)
Bcip nbt alp color development substrate, by Beyotime (2 mentions)
Calcium assay kit, by Nanjing Jiancheng (2 mentions)
Triton x 100, by Merck Group (2 mentions)
128 protocols using alp assay kit
1
Serum and Bone ALP Activity Analysis
2
Quantifying Osteoblast ALP Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After treatment with OM for 1, 3, 5 and 7 days, MC3T3-E1 cells were rinsed with PBS, solubilized in lysis buffer (50 mM Tris, 100 mM glycine) for 5 min and centrifuged (2,000 × g; room temperature; 5 min) to collect the supernatant. The ALP activity in the supernatant was then determined using ALP Assay kits (Nanjing Jiancheng Bioengineering Institute) according to the manufacturer's instructions. The ALP activity was normalized to total the protein concentration, which was determined using BCA Protein Assay kits (Thermo Fisher Scientific, Inc.).
3
Serum Biomarker Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Serum was thawed and analyzed for the Ca concentration using a microplate reader with Ca assay kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China); serum phosphorus concentration was determined using the molybdenum blue method according to Goldenberg and Fernandez [30 (link)]; serum intact FGF23 concentration was assessed using an ELISA kit specially used for poultry according to the manufacturer’s protocol (Nanjing Jiancheng Bioengineering Institute, Nanjing, China); serum PTH level was measured using an ELISA kit according to the manufacturer’s protocol (Nanjing Jiancheng Bioengineering Institute, Nanjing, China); and serum 1,25(OH)2D3 level was measured by an ELISA kit by Beijing Sino-UK Institute of Biological Technology. The ALP activities in serum and tibia were measured using a microplate reader with ALP assay kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). The tibia BGP concentration was determined by the method of ELISA with a commercial kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China).
4
Quantifying Bone Mineralization Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The ALP activity was measured using a microplate reader with ALP assay kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). The BGP content was determined by the method of ELISA with a commercial kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer's instructions. The ALP activity and the BGP content in each sample were normalized by the protein concentration.
5
Quantitative Evaluation of Osteogenic Differentiation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ALP staining was conducted as described in the section ‘BMSC culture and identification’. To evaluate ALP activity, osteoblasts on samples were lysed with RIPA lysis buffer (Beyotime, China) and were assessed using an ALP assay kit (Jiancheng Inc, China) and a BCA protein assay kit (Beyotime, China) according to the manufacturer’s instructions. ALP activity was normalized to the total protein content of the cell lysate.
Alizarin red S staining was conducted as described in the section ‘BMSC culture and identification’. The stains were eluted with 10% cetylpyridinium chloride in 10 mM sodium phosphate (pH 7.0) for quantification analysis.
Alizarin red S staining was conducted as described in the section ‘BMSC culture and identification’. The stains were eluted with 10% cetylpyridinium chloride in 10 mM sodium phosphate (pH 7.0) for quantification analysis.
6
Quantitative Analysis of Osteogenic ALP
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After osteogenic induction for 7 days, cells were subjected to ALP staining and quantification as previously described [22 (link)]. ALP staining was conducted using the NBT/BCIP staining kit (CoWin Biotech, Beijng, China). An ALP assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) was used to quantify ALP activity. The bicinchoninic acid (BCA) method with a Pierce protein assay kit (Thermo Scientific, Rockford, IL, USA) was used to measure the total protein concentration. After normalization to the total protein concentration, the ALP levels relative to the control group were analyzed.
7
Osteogenic Differentiation of MC3T3-E1 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MC3T3-E1 cells were incubated in a 24-well plate at a density of 2×104 cells/well (n = 4). After culturing for 24 h, the medium was replaced with fresh osteogenic induction medium containing dexamethasone (100 nM, Sigma, United States), β-glycerophosphate (10 mM, Sigma, United States) and L-ascorbic acid (50 mM, Sigma, United States). After 4 and 7 d of osteogenic induction, the total amount of protein was examined by a BCA protein assay kit (Beyotime, China). An ALP assay kit (Nanjing Jiancheng, China) was employed for the quantitative assessment of ALP. Finally, the ALP activity was measured according to the instructions, and it was standardized to the total protein content. Afterwards, at testing time points on 4 and 7 d of induction, the cells were stained with a BCIP/NBT Kit (Beyotime, China) and observed by stereomicroscopy (Olympus, Japan).
8
Silencing lncRNA SMILR Enhances Osteogenesis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DPSCs were transfected with siRNA (Sense 5′-CCUAAACUCGAAUCUGAUUTT - 3′, Antisense 5′-AAUCAGAUUCGAGUUUAGGTT - 3′) to reduce the expression of lncRNA SMILR using the Micropoly-transfecter Cell Reagent (Micropoly, Xian, Shanxi, China). DPSCs transfected with non-target siRNA (Sense 5′- UUCUCCGAACGUGUCACGUTT- 3′, Antisense 5′- ACUUGACACGUUCGGAGAATT- 3′) were used as control group. All DPSCs were cultured in the osteogenic medium for 7 days (DPSCs were transfected again on the fourth day to enhance the effect of siRNA), then were collected for Alkaline phosphatase (ALP) staining (by NBT/BCIP staining kit (Beyotime, Shanghai, China)), ALP activity assay (by ALP assay kit (Jiancheng, Nanjing, China)) and qRT-PCR experiments.
9
Alkaline Phosphatase Activity in MC3T3-E1 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After culturing MC3T3-E1 cells (2 × 104 cells/cm2) on various samples for 3 and 7 d, an ALP Assay Kit (Nanjing Jiancheng Co., China) was used to determine the level of ALP activity. Briefly, 1% Triton-100 was added to lyse cells for 40 min after washing the samples in PBS. The substrates, deionized water for the blank, standard solution for the control, and corresponding cell lysates for samples were added to 96-well plates, respectively. They were then mixed with buffer solution and Matrix fluid before incubation at 37°C for 15 min. Chromogenic agent was then added and the OD values were measured at 520 nm. The protein content of each sample was also measured using a BCA Protein Assay Kit (Beyotime, China).
10
Alkaline Phosphatase Activity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
On day 14, ALP activity was measured using an ALP assay kit (Jiancheng Corp, Nanjing, China) according to the manufacturer's protocols. The enzyme activity was expressed as micromoles of reaction product per minute per total protein.
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!