Pre-osteoblast MC3T3-E1 cells were cultured in an osteogenic medium (complete DMEM supplemented with 50 μg/mL ascorbic acid and 5 mM β-glycerophosphate).[34 (link)] To evaluate the model of differentiated MC3T3-E1 cells, total alkaline phosphatase activity and calcified nodules were determined at days 1, 7, and 14 of culture. Enzymatic activity was determined by spectrophotometry (405 nm) using a commercial kit (ALP 405 AA Wiener lab) and expressed concerning the total protein content assessed by spectrophotometry (540 nm; commercial kit Proti U/LCR Wiener Lab). Protein assessment was performed in a homogenization obtained by washing the cells with phosphate-buffered saline, adding radioimmunoprecipitation assay buffer and using a scrapper and sonication to break the cell walls. The mineralization assay was performed by fixing the cells with 4% formaldehyde for 10 min and staining them with a 2% alizarin red (Sigma-Aldrich) solution for 30 min at room temperature, allowing the visualization of calcified nodules under a microscope.[27 (link)]
Proti u lcr
Manufactured by Wiener Lab
Sourced in Argentina
About the product
The Proti U/LCR is a laboratory instrument designed to perform quantitative protein analysis. It utilizes a combination of UV absorbance and luminescent detection techniques to accurately measure the concentration of proteins in a sample.
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9 protocols using «proti u lcr»
1
Osteoblast Differentiation and Mineralization
2
Quantitative Biochemical Analysis of Renal Function
Creatinine was determined by the alkaline picrate colorimetric method in plasma and urine samples. The total urinary protein was determined with a quantitative colorimetric method based on the use of pyrogallol red (Proti U/LCR, Wiener lab, Rosario, Argentina). UAE was measured with a competitive chemiluminescent enzyme immunoassay (Immulite/Immulite 1000 Albumin, DPC, Los Angeles, CA, USA). Creatinine, total protein, and albumin determination were carried out as previously described [15 (link),16 (link)].
3
Renal Function Assessment Protocol
Blood was immediately centrifuged to separate plasma for creatinine, Na + , and K + measurements. Urine volume was measured gravimetrically. Urinary creatinine, Na + , K + , and protein were measured. GFR was estimated by creatinine clearance and was referred to 100 g bw. Creatinine measure was determined using a commercial kit (Creatinina, Wiener Lab, Rosario, Argentina). Na + and K + measurements were performed by an electrolyte analyzer with ion-selective electrodes (Roche Diagnostics, Switzerland). Urinary protein concentration was determined by a commercial kit based on pyrogallol red-molybdate (Proti U/LCR, Wiener Lab, Rosario, Argentina).
Fractional excretion of sodium (FE Na ), and potassium (FE K ) were calculated by relation between sodium and potassium clearance and GFR. Fractional excretion of water (FE H2O ) was calculated by relation between plasmatic and urinary creatinine. Urine protein excretion load (E Protein ) was also calculated.
Fractional excretion of sodium (FE Na ), and potassium (FE K ) were calculated by relation between sodium and potassium clearance and GFR. Fractional excretion of water (FE H2O ) was calculated by relation between plasmatic and urinary creatinine. Urine protein excretion load (E Protein ) was also calculated.
4
Urine Analysis in Metabolic Cages
Twenty four-hour urine samples were collected using metabolic cages. Animals were allowed to acclimatize to metabolic cages for two days and then fasted for 24 h before the collection of urine. Urine samples were analyzed for total protein using a kit provided by Wiener (Proti U/LCR; Wiener Lab., Rosario, Argentina). Urinary and plasmatic sodium and potassium concentrations were evaluated using an ion analyzer (Tecnolab; Mod T-412). Kinetic determinations of serum and urinary creatinine concentrations were evaluated using a kit provided by Wiener (Wiener Lab., Rosario, Argentina). Urine volume was measured gravimetrically.
5
Rat Urine Protein Quantification
The urine of rats were recollected in a metabolic cage for 24 h and the protein contents were quantified while using the pyrogallol red molybdate method (Proti U/LCR, Wiener Lab, Rosario, Argentina).
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