Nephelostar nephelometer

Manufactured by BMG Labtech

Sourced in Germany

The NEPHELOstar is a nephelometer instrument designed for measuring light scattering in liquid samples. It is used to quantify the concentration of particles or molecules in a solution.

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Lab products found in correlation

Dynapro plate reader 2, by Wyatt Technology (1 mentions) Colorimetric assay, by Merck Group (1 mentions) Fetuin a, by R&D Systems (1 mentions) Live dead baclight bacterial viability kit, by Thermo Fisher Scientific (1 mentions) Costar plate, by Corning (1 mentions) Turbidity 4000 ntu calibration standard formazin, by Merck Group (1 mentions) Freedom evo 100, by Tecan (1 mentions)

Spelling variants of this product

NEPHELOstar (16 citations) Nephelostar Plus nephelometer (3 citations)

7 protocols using nephelostar nephelometer

Serum Biomarkers in Dialysis Patients

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Sera from both the HD and HDF patients were drawn before and after dialysis during all in centre dialysis sessions in one week. Sera were analysed for Ca, magnesium, albumin, P, bicarbonate and CRP on a routine automated analyser (Cobas c502 immunochemistry analyser (Roche Diagnostics, Almere, The Netherlands). For the analysis of fetuin-A and the CPP transformation time (T₅₀) an additional 5ml, non-additive BD-vacutainer glass serum tube was collected. Within 240 minutes after collection these samples were centrifuged at room temperature (20°C) for 15 minutes. Of this 800 ul of serum was stored at 4°C and send to the lab of Calciscon AG in Bern, Switzerland, where they were analysed within 72 hours after collection.
The serum fetuin-A concentration was measured by nephelometry, a test first established by the Jahnen-Dechent group [28 (link)]. T₅₀ was determined by the method described by Pasch et al. [13 (link)]. Samples were supersaturated by adding Ca (10mM) and P (6mM) to initiate the formation of primary CPP. The time of spontaneous transformation to secondary CPPs was measured by a nephelostar nephelometer (BMG Labtech, Ortenberg, Germany).
Correction for hemoconcentration for fetuin-A was performed by the method of Bergstrom et al. [29 (link)]: uncorrected post dialysis fetuin-A (g/L)/(1+ (Δ body weight (kg)/0.2*post dialysis body weight (kg)).

Dekker M., Pasch A., van der Sande F., Konings C., Bachtler M., Dionisi M., Meier M., Kooman J, & Canaud B. (2016). High-Flux Hemodialysis and High-Volume Hemodiafiltration Improve Serum Calcification Propensity. PLoS ONE, 11(4), e0151508.

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Calcification Propensity Test Protocol

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T 50 -time was determined by Calciscon AG, Bern, Switzerland as described previously [15] . In brief, the calcification propensity test is an in vitro nano-particle assay, which measures the time of conversion of primary to secondary CPPs. Serum samples were supersaturated with calcium and phosphate to increase their final concentrations by 10 and 6 mM, respectively. Secondary CPP formation time (T 50 -time) was measured by a nephelostar nephelometer (BMG Labtech, Ortenberg, Germany). Other parameters were measured as detailed in Gaggl et al. [25] .
The

Aigner C., Cejka D., Sliber C., Fraunschiel M., Sunder-Plassmann G, & Gaggl M. (2019). Oral Sodium Bicarbonate Supplementation Does Not Affect Serum Calcification Propensity in Patients with Chronic Kidney Disease and Chronic Metabolic Acidosis. Kidney & blood pressure research, 44(2).

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Solubility Determination of Compound 45 and Carnosine

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Example 4

The solubility of Compound 45 and carnosine in physiologic buffer (phosphate buffered saline, pH 7.4; PBS) were determined using a BMG Labtech NEPHELOstar nephelometer (Offenburg, Germany) with Galaxy analytical software, version 4.32R2. Stock solutions of each compound were prepared initially in dimethyl sulfoxide (DMSO); Compound 45 was soluble at a concentration of 50 mM in DMSO, while carnosine was not. Subsequently, dilution series of both compounds were prepared directly in PBS (pH 7.4) for evaluation of solubility using nephlometry, with a top concentration of 500 mg/mL for each compound and serial 1:2 dilutions. A positive control compound was included to qualitatively illustrate concentration-solubility relationships using heat maps. Compound 45 was soluble in physiologic buffer at a concentration of 500 mg/mL, while carnosine was soluble only at concentrations of 250 mg/mL.

US10028935B2. Stabilized multi-functional antioxidant compounds and methods of use (2018-07-24). XPD Holdings, LLC [US]. Inventors: Marc Bailie [US], Steven K. Duddy [US], Jim Herman [US].

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Measuring Serum Biomarkers in Mineralization

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Serum T50 was measured by Calciscon AG, Biel, Switzerland, as previously described using a Nephelostar nephelometer (BMG Labtech, Ortenberg, Germany)^{40 (link)}. The mean interassay CV_A for T50 was 3.4%. CPP-II hydrodynamic radius was measured by dynamic light scattering using a DynaPro Plate Reader II (Wyatt Technology, Santa Barbara, CA, USA) as described by Chen et al.^{47 (link)} The interassay CV_A for CPP-II size was 4%. Commercial immunoassays were used to measure iFGF23 (Kainos Laboratories, Tokyo, Japan), 1,25 dihydroxyvitamin D Immunodiagnostic Systems, Boldon, UK), and fetuin-A (R&D Systems, Minneapolis, USA) according to the manufacturer’s instructions. Mean interassay CV_A were 3.8%, 5.5%, and 3.2%, respectively. Serum citrate was measured using a colorimetric assay (Sigma-Aldrich, Darmstadt, Germany) with a mean interassay CV_A of 3.5%.

Tiong M.K., Cai M.M., Toussaint N.D., Tan S.J., Pasch A, & Smith E.R. (2022). Effect of nutritional calcium and phosphate loading on calciprotein particle kinetics in adults with normal and impaired kidney function. Scientific Reports, 12, 7358.

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Vorin-cat Expression in E. coli

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E. coli BL21(DE3) cells were transformed with 100 ng of the appropriate expression vector or combination of vectors by heat shock method, and a single colony was used to inoculate a 5 mL LB culture containing either 30 µg/mL kanamycin or 30 µg/mL kanamycin and 100 µg/mL streptomycin. The 5 mL cultures were incubated at 37 °C for 6 h with shaking. Culture OD₆₀₀ readings were taken after 18 h, and the values were used to dilute each culture to OD₆₀₀ = 2 × 10⁻⁴ in sterile 1.5 mL tubes with fresh LB containing the appropriate antibiotic. Expression of Vorin-cat and VorinI was induced by addition of IPTG to a final concentration of 0.1 mM. Each control or experimental culture was dispensed into a 96-well plate in 200 µL aliquots. Plates were incubated at 35°C with shaking using a NEPHELOstar nephelometer (BMG LABTECH) and culture turbidity was measured in nephelometric turbidity units (NTU) every 5 min for 16.5 h. The BacLight Live/Dead bacterial viability kit (L-7012, Molecular Probes) was used to assess cell viability after a three-hour induction of vorin-cat in E. coli Lemo21 cells.

Tremblay O., Thow Z, & Merrill A.R. (2020). Several New Putative Bacterial ADP-Ribosyltransferase Toxins Are Revealed from In Silico Data Mining, Including the Novel Toxin Vorin, Encoded by the Fire Blight Pathogen Erwinia amylovora. Toxins, 12(12), 792.

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Nephelometric Turbidity Measurement Protocol

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Solubility measurement samples were prepared in a 96-well Corning Costar plate with a total sample volume of 200 µL in either PBS buffer with 1% DMSO (v/v) or pure water. A BMG Labtech NEPHELOstar nephelometer was used to measure turbidity of the samples with the following parameters per the instrument manufacturer: Gain = 80, Laser Focus= 2.50 mm, and Laser Intensity = 90%. Samples were shaken in the nephelometer by orbital shaking for 3 s prior to the turbidity measurement. Sample values were blanked in the instrument software using the corresponding PBS and DMSO condition values. Turbidity 4000 NTU Calibration Standard Formazin (Millipore-Sigma) was used as a positive control for all turbidity measurements. The turbidity of each sample was measured within 20 min of initial preparation.

Liu H., Iketani S., Zask A., Khanizeman N., Bednarova E., Forouhar F., Fowler B., Hong S.J., Mohri H., Nair M.S., Huang Y., Tay N.E., Lee S., Karan C., Resnick S.J., Quinn C., Li W., Shion H., Xia X., Daniels J.D., Bartolo-Cruz M., Farina M., Rajbhandari P., Jurtschenko C., Lauber M.A., McDonald T., Stokes M.E., Hurst B.L., Rovis T., Chavez A., Ho D.D, & Stockwell B.R. (2022). Development of optimized drug-like small molecule inhibitors of the SARS-CoV-2 3CL protease for treatment of COVID-19. Nature Communications, 13, 1891.

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Serum T50 Calcification Propensity Measurement

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Serum samples for T50 measurement were obtained from the participants through venipuncture at the same moment when blood sampling for routine blood chemistry and PPi was performed. Serum samples were stored at −80 °C without thawing. T50 calcification propensity measurements were performed at Calciscon AG, Nidau, Switzerland, as previously described [10 (link)]. In short, serum samples were thawed, vortexed and centrifuged after which they were supersaturated with calcium and phosphate solutions (pH 7.40). Pipetting was performed using an automated high-precision pipetting system (Freedom EVO 100, Tecan, Switzerland). Samples were then measured in triplicate in 96-well plates at 37 °C for 600 min in a Nephelostar nephelometer (BMG Labtech, Ortenberg, Germany), hence, live-monitoring the transformation of primary to secondary CPPs in the different samples (batch 1: n = 23; batch 2: n = 34). To determine the half-maximal transformation time (T50), data analyses of nonlinear regression curves were performed using the Calciscon T50 Analysis Software. Intra- and interassay coefficients of variation were 2.2% and 3.4%, respectively.

Nollet L., Van Gils M., Fischer S., Campens L., Karthik S., Pasch A., De Zaeytijd J., Leroy B.P., Devos D., De Backer T., Coucke P.J, & Vanakker O.M. (2022). Serum Calcification Propensity T50 Associates with Disease Severity in Patients with Pseudoxanthoma Elasticum. Journal of Clinical Medicine, 11(13), 3727.

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