Lambda 25 uv vis spectrophotometer

Manufactured by PerkinElmer

Sourced in United States, Italy

The Lambda 25 UV-Vis spectrophotometer is a compact, high-performance instrument designed for accurate and reliable absorbance measurements in the ultraviolet and visible light spectrum. It features a dual-beam optical system and a broad wavelength range for a variety of analytical applications.

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Spelling variants of this product

Lambda 25 (458 citations) Lambda 25 spectrophotometer (140 citations) UV-Vis spectrophotometer (137 citations) Lambda 25 UV/VIS (73 citations) UV spectrophotometer (30 citations) Lambda 25 UV spectrophotometer (10 citations) Lambda 25 UV (9 citations) Lambda 25 UV/vis absorption spectrophotometer (4 citations)

151 protocols using lambda 25 uv vis spectrophotometer

Antioxidant Enzyme Activities and Glutathione Levels

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Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR) were assessed in erythrocytes, adipose tissue, and liver. The total SOD and CAT activities were measured according to the methods developed by Mirsa and Fridovich [27 (link)] and Cohen et al. [28 (link)], respectively, using a Lambda 25 UV-Vis spectrophotometer (Perkin Elmer, Shelton, CT, USA). GPx and GR activities were measured according to the method developed by Wheeler et al. [29 (link)] using a COBAS MIRA autoanalyzer (Roche Diagnostics System, Madrid, Spain).
Reduced glutathione (GSH) and oxidized glutathione (GSSG) in plasma, erythrocytes, adipose tissue, and liver were measured according to the method developed by Hissin and Hilf [30 (link)] using an LS55 fluorescence spectrophotometer (Perkin Elmer, Shelton, CT, USA) and the corresponding standard curves (Sigma-Aldrich, Madrid, Spain). In addition, the GSSG/GSH ratio was calculated as a biomarker of the redox state.
Measurements in erythrocyte samples were normalized to hemoglobin (Hb) concentration in total blood. Hb concentration was measured according to the Drabkin method [31 (link)] using a Lambda 25 UV-Vis spectrophotometer (Perkin Elmer, Shelton, CT, USA), and an Hb standard (Spinreact, Girona, Spain).

Miralles-Pérez B., Nogués M.R., Sánchez-Martos V., Taltavull N., Méndez L., Medina I., Ramos-Romero S., Torres J.L, & Romeu M. (2021). The Effects of the Combination of Buckwheat D-Fagomine and Fish Omega-3 Fatty Acids on Oxidative Stress and Related Risk Factors in Pre-Obese Rats. Foods, 10(2), 332.

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Phenolic, Anthocyanin, and Flavonoid Quantification

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Phenolic compounds content was determined according to the method proposed by Bizzo et al. (2014) (link). For anthocyanin determination, plant samples (50 mg of FW) were incubated with 5 mL of methanol:HCl (99:1) and kept in the dark for 24 h. The extract was then centrifuged at 13,000 x g for 10 min. The absorbance of the supernatant was measured at 550 nm using a Lambda 25 UV-vis spectrophotometer (Perkin Elmer, USA). Anthocyanin concentration (nmol g -1 FW) was calculated using an extinction coefficient of 33,000 mol -1 cm -1 (Wanger, 1979) (link).
For flavonoid content determination, 50 mg of FW were mixed with 5 mL of ethanol:acetic acid (99:1). Then, the samples were gently boiled for 10 min in a water bath at 80 °C. The absorbance was measured at 330 nm using a UV-vis Lambda 25 spectrophotometer (Perkin Elmer, USA). Flavonoid concentration was expressed as absorbance units (AU 330 ) g -1 FW.

Castillo Loría K., Emiliani J., Bergara C.D., Herrero M.S., Salvatierra L.M, & Pérez L.M. (2019). Effect of daily exposure to Pb-contaminated water on Salvinia biloba physiology and phytoremediation performance. Aquatic toxicology (Amsterdam, Netherlands), 210.

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Rapid PANI Film Fabrication

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PANI patches (n = 4) were fabricated as described above, except for the polymerization time, which was decreased from 3 hours to 5 min to obtain films with reduced opacity, allowing for the transmission of the UV beam. Samples were prepared on glass slides (4 cm × 1 cm) and incubated in a 3-ml cuvette containing PBS. Their absorption spectra were recorded at predetermined time points with a PerkinElmer Lambda 25 UV/Vis spectrophotometer in the range between 300 and 900 nm.

Mawad D., Mansfield C., Lauto A., Perbellini F., Nelson G.W., Tonkin J., Bello S.O., Carrad D.J., Micolich A.P., Mahat M.M., Furman J., Payne D., Lyon A.R., Gooding J.J., Harding S.E., Terracciano C.M, & Stevens M.M. (2016). A conducting polymer with enhanced electronic stability applied in cardiac models. Science Advances, 2(11), e1601007.

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Glucose-Oxidase Activity Determination Protocol

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The glucose-oxidase activity was determined according to Cohen [39 (link)] and Sagona et al. [40 (link)] with some modifications. For each protein extract, 1.5 mL of solution containing 100 mM Hepes buffer pH7, 0.1 mM EDTA, 5 mM D-glucose, Diaminobenzidine (DAB) (0.18 mg/mL), and horseradish peroxidase (HRP) (0.02 mg/mL) was prepared. Absorbance was obtained at a wavelength of λ = 352 nm at times 0 and 120 min. All enzymatic analyses were conducted using a Lambda 25 UV/VIS spectrophotometer (Perkin Elmer, Milano, Italy).

Sagona S., Coppola F., Giannaccini G., Betti L., Palego L., Tafi E., Casini L., Piana L., Dall’Olio R, & Felicioli A. (2022). Impact of Different Storage Temperature on the Enzymatic Activity of Apis mellifera Royal Jelly. Foods, 11(20), 3165.

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Quantifying Total Phenolic Content

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The extract was determined for total phenolic content (TPC) using the Folin-Ciocalteu method with minor modifications^{58 (link)}. The freeze-dried extract was dissolved in distilled water to a concentration of 1 mg/mL. Gallic acid was used as the standard and a calibration curve was established (0 – 1000 mg/mL). The extract or gallic acid (0.5 mL) was added to 2.5 mL Folin-Ciocalteu reagent (tenfold diluted with distilled water) and mixed thoroughly for 3 min. Sodium carbonate (2 mL, 7.5% w/v) was added to the mixture and the mixture was allowed to stand for 30 min at room temperature. The absorbance of the mixture was measured using a Perkin Elmer Lambda 25 UV–VIS spectrophotometer (Norwalk, USA) at 760 nm. TPC was expressed as mg gallic acid equivalent/g dry weight of extract (mg GAE/g extract).

Jeyaraj E.J., Lim Y.Y, & Choo W.S. (2022). Antioxidant, cytotoxic, and antibacterial activities of Clitoria ternatea flower extracts and anthocyanin-rich fraction. Scientific Reports, 12, 14890.

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Characterizing Fluorescent Protein Optical Properties

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Absorption spectra and excitation/emission spectra of FPs were measured separately using a LAMBDA-25 UV/Vis spectrophotometer (Perkin Elmer, Waltham, USA) and a LS55 fluorescence spectrophotometer (Perkin Elmer, Waltham, USA) equipped with a red sensitive photomultiplier tube (Hamamatsu Photonics, Hamamatsu, Japan). Molar extinction coefficients were quantified using a alkali denaturation method [12 ,13 ]. In order to measure accurately the quantum yields, according to the description of optical dilution measurement [14 ], protein samples were first diluted in PBS (pH7.4) with UV absorption value in the range of 0.01–0.05. Fluorescence intensity of the mutant FP was compared with that of mNeptune (a quantum yield of 0.2). In order to facilitate the brightness comparisons between different FP variants, the brightness of all the proteins is converted to the relative brightness, which is relative to the brightness of EGFP (the product of quantum yield and extinction coefficient compared to the brightness of EGFP (53,000M^-1cm^-1×0.60) [15 (link)]).

Li Z., Zhang Z., Bi L., Cui Z., Deng J., Wang D, & Zhang X.E. (2016). Mutagenesis of mNeptune Red-Shifts Emission Spectrum to 681-685 nm. PLoS ONE, 11(4), e0148749.

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Liver Gluconeogenic and Glycolytic Enzymes

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The activities of gluconeogenic enzymes pyruvate carboxylase (PC, EC 6.4.1.1), phosphoenolpyruvate carboxykinase (PEPCK, EC 4.1.1.32), fructose-1,6-bisphosphatase (FBPase, EC 3.1.3.11) and glucose-6-phosphatase (G6Pase, EC 3.1.3.9) were measured in liver, as previously described [14 (link)]. The activities of the glycolytic enzymes glucokinase (GK, EC 2.7.1.1), phosphofructokinase-1 (PFK1, EC 2.7.1.11) and pyruvate kinase (PK, EC 2.7.1.40) were also measured in liver as previously described [15 (link)]. All assays were performed using a Perkin Elmer Lambda 25 UV/Vis spectrophotometer equipped with a Peltier heating control system and 9 cell changer (Perkin Elmer, Shelton, CT). Reactions were followed at 340 nm (ε=6.22 mM⁻¹.cm⁻¹) for GK, PFK1, PK, FBPase and PEPCK, 412 nm (ε=13.6 mM⁻¹.cm⁻¹) for PC and 510 (ε=6.66 mM⁻¹.cm⁻¹) for G6Pase. All enzyme activities were expressed as μmol/min/mg protein, and presented as mean±SEM determined from at least six animals.

Hagopian K., Kim K., López-Dominguez J.A., Tomilov A.A., Cortopassi G.A, & Ramsey J.J. (2016). Mice with low levels of Shc proteins display reduced glycolytic and increased gluconeogenic activities in liver. Biochemistry and Biophysics Reports, 7, 273-286.

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Quantifying Lipid Peroxidation in Wheat Seedlings

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Lipid peroxidation levels in wheat seedlings were estimated by measuring the thiobarbituric acid reactive substances (TBARS), according to the method described by Verma and Dubey [57 (link)]. This method is based on the formation of red pigment, generated by the reaction of lipid peroxidation breakdown products like malondialdehyde (MDA) with thiobarbituric acid at an optimum pH of 3.5. Briefly, the frozen wheat powder was homogenized with 0.1% trichloroacetic acid (TCA) solution (1/5, w/v) and centrifuged for 10 min at 10,000× g and 4 °C. The reaction mixture that consisted of 0.5 mL of tissue extract and 1 mL of reagent (0.5% thiobarbituric acid in 20% TCA) was incubated for 30 min at 95 °C on a TS-100 Thermo-Shaker (Biosan, Riga, Latvia). After cooling the reaction mixture, the produced red pigment was measured at 532 and 600 nm on a LAMBDA 25 UV-Vis spectrophotometer equipped with UV WinLab v6.0.4 software package (PerkinElmer, Waltham, MA, USA). The results were expressed as nmol/g of FW.

Vuković R., Čamagajevac I.Š., Vuković A., Šunić K., Begović L., Mlinarić S., Sekulić R., Sabo N, & Španić V. (2022). Physiological, Biochemical and Molecular Response of Different Winter Wheat Varieties under Drought Stress at Germination and Seedling Growth Stage. Antioxidants, 11(4), 693.

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Dialysis-Related Biochemical Markers

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Six milliliters of venous blood sample just before commencing dialysis (pre-HD) and at the termination of dialysis (post-HD) was collected in additive free tubes. The blood samples were allowed to stand for 30 min, centrifuged at 3000 rpm for 15 min and the separated serum was stored at −80 °C until further analysis. Serum urea, creatinine, uric acid, total cholesterol, triglycerides, HDL, calcium, phosphorus, total protein, and albumin were estimated using commercial kits. Low density lipoprotein (LDL) and very low density lipoprotein (VLDL) were calculated by Freidewald’s equation [15 (link)]. hs-CRP was estimated by immunoturbidimetry method by commercial kits. All the above parameters were analyzed on clinical chemistry Auto analyzer Beckman Coulter DXC 600 Synchron (‎Brea, CA). Plasma MDA and ferric reducing ability of plasma (FRAP) were estimated by spectrophotometric method using Perkin-Elmer spectrophotometer, Lambda 25 UV/VIS Spectrophotometer (Waltham, MA) [16 (link),17 (link)]. Pentraxin-3 was estimated by ELISA method using the commercially available human Pentraxin ELISA kit (Hycult, Beutelsbach, Germany).

Sangeetha Lakshmi B., Harini Devi N., Suchitra M.M., Srinivasa Rao P.V, & Siva Kumar V. (2018). Changes in the inflammatory and oxidative stress markers during a single hemodialysis session in patients with chronic kidney disease. Renal Failure, 40(1), 534-540.

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Cultivation of Diverse Diatom Species

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Thalassiosira pseudonana Hasle & Heim., strain CCMP1335 subcultured in the lab since 2013, Phaeodactylum tricornutum Bohlin, strain CCMP2561, and Navicula pelliculosa (Brébisson ex Kützing) Hilse, strain CCAP 1050/9, were grown in artificial sea water enriched with Guillard’s “F/2” nutrients plus silicon (F/2+Si) as described previously [34 (link)]. The diatom N. pelliculosa was also cultured in a freshwater medium in the absence of NaCl and KCl, and NaH₂PO₄·2H₂O (0.036 mM) was replaced by KH₂PO₄ (0.036 mM). Asterionella formosa Hassal, isolated from the English Lake District (Esthwaite Water; strain BG1), was grown in Diatom Medium (DM) as described [19 (link)]. Cultures were maintained at 18 °C in a growth cabinet (Innova 4230; New Brunswick Scientific, Edison, NJ, USA) with continuous light at 50 µmol photon m⁻² s⁻¹, constantly shaken at 90 rpm, and bubbled with air containing either 400 ppm or 20,000 ppm CO₂ at a gas flow rate of 320 mL min⁻¹ L⁻¹ of culture. For growth curves, optical density was followed at 750 nm using a Perkin Elmer Lambda 25 UV/VIS spectrophotometer (Waltham, MA, USA).

Jensen E.L., Yangüez K., Carrière F, & Gontero B. (2019). Storage Compound Accumulation in Diatoms as Response to Elevated CO2 Concentration. Biology, 9(1), 5.

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