Enzymatic kit

Manufactured by Fujifilm

Sourced in Japan, United States, Germany

The Enzymatic kit is a laboratory product designed to aid in the detection and quantification of specific enzymes. It provides a standardized and reliable method for measuring enzyme activity in various samples. The kit contains the necessary reagents and protocols to perform the analysis.

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Lab products found in correlation

Superdex 200, by GE Healthcare (1 mentions) Tubulin 2144, by Cell Signaling Technology (1 mentions) Ng nitro l arginine methyl ester l name, by Merck Group (1 mentions) Gw6471, by Cayman Chemical (1 mentions) Pemafibrate, by Kowa (1 mentions) Amplex red cholesterol assay kit, by Thermo Fisher Scientific (1 mentions) Tr0100 kit, by Merck Group (1 mentions) Whatman, by Cytiva (1 mentions) Unicel dxc 600, by Beckman Coulter (1 mentions) Superose 6 10 300 gl column, by GE Healthcare (1 mentions) Griess reagent kit, by Thermo Fisher Scientific (1 mentions) Infinity triglycerides reagent, by Thermo Fisher Scientific (1 mentions) Milliplex kit, by Merck Group (1 mentions) Enzymatic kits, by Eiken Chemical (1 mentions) Apo ai, by Eiken Chemical (1 mentions) Enzymatic kit, by Roche (1 mentions) F2hfhsd, by Oriental Yeast (1 mentions) C57bl 6j, by Jackson ImmunoResearch (1 mentions) Adiponectin, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Enzymatic assay kits (34 citations) Enzymatic colorimetric kits (11 citations) Enzymatic colorimetric assay kit (9 citations) Wako enzymatic kit (5 citations) Cholesterol E Enzymatic Kit (3 citations) Enzymatic colorimetry assay kits (2 citations) Enzymatic reagent kit (2 citations) Enzymatic cholesterol kit (2 citations)

36 protocols using enzymatic kit

Quantifying Plasma Lipid Profiles

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Plasma concentrations of total cholesterol (C) and TAG were measured with enzymatic kits no. 61219 for C and no. 61236 for TAG (both from bioMérieux SA, Geneva, Switzerland). PL and free cholesterol (FC) concentrations were measured using enzymatic kits from Wako Pure Chemical Industries Ltd., Osaka, Japan (no. 296–63801 for PL; no. 435–35801 for FC). High density lipoprotein-cholesterol (HDL-C) was measured using an immunoinhibition method (kit no. 412–72395) and LDL-C with an enzymatic kit (no. 419–24017) (both from Wako Chemical GmbH, Neuss, Germany).
Concentrations of cholesteryl esters (CE) and VLDL-C were calculated according the following equations: [7 (link)]

Cholesteryl esters = cholesterol - free cholesterol;

VLDL-C = TAG / 5

Gross J.J., Kessler E.C., Albrecht C, & Bruckmaier R.M. (2015). Response of the Cholesterol Metabolism to a Negative Energy Balance in Dairy Cows Depends on the Lactational Stage. PLoS ONE, 10(6), e0121956.

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Plasma Metabolic Biomarker Analysis

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Blood samples were collected through arterial catheters after a 6 h fast unless otherwise noted. Blood was immediately placed in heparinized‐lithium tubes, separated by centrifugation, and stored at 4°C or −80°C for long‐term storage. Plasma glucose concentrations were measured using the YSI Biochemistry Analyzer or Ascensia Elite glucose meter (survival studies; Martin‐Montalvo et al., 2013 (link)); plasma insulin was measured by RIA or ELISA. Plasma NEFAs, total cholesterol, and HDL‐C concentrations were measured using enzymatic kits from Wako according to the manufacturer's instructions. Plasma triglycerides were measured using the Sekisui Triglyceride‐SL Kit (Sekisui Diagnostics) and plasma βOHB, AST, ALT, and BUN by COBAS (Roche Diagnostics).

Goedeke L., Murt K.N., Di Francesco A., Camporez J.P., Nasiri A.R., Wang Y., Zhang X., Cline G.W., de Cabo R, & Shulman G.I. (2022). Sex‐ and strain‐specific effects of mitochondrial uncoupling on age‐related metabolic diseases in high‐fat diet‐fed mice. Aging Cell, 21(2), e13539.

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Blood Lipid Profile Analysis

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Whole blood was collected from the vena cava of overnight‐fasted mice. Serum was separated and total cholesterol, HDL, and LDL were analyzed using enzymatic kits from Wako Diagnostics, triglycerides using Infinity^™ Triglycerides Reagent and nonesterified free fatty acids using an enzymatic kit from Zenbio (Research Triangle Park, Durham, NC).

Hammond C.L., Wheeler S.G., Ballatori N, & Hinkle P.M. (2015). Ostα−/− mice are not protected from western diet‐induced weight gain. Physiological Reports, 3(1), e12263.

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HDL Fractionation and Characterization

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HDL particles were fractionated as described [17 (link)]. Briefly, 450 μL of pooled plasma was applied to a Superdex 200 gel filtration column (10/300 GL, GE Healthcare) in an ÄKTA fast protein liquid chromatography (FPLC) system, and separated at a flow rate of 0.3 mL/min in standard Tris buffer (10 mM Tris, 0.15 M NaCl, 1 mM EDTA). 48 fractions were collected (GE Healthcare) and stored at 4 °C.
FPLC fractions were tested for lipid and total protein concentrations by using Colorimetric kits and bicinchoninic acid (BCA). Protein content of lipoprotein fractions were quantified by the absorbance at OD280. Total cholesterol (TC) and triglyceride (TG) levels were assayed by Colorimetric kits. Phospholipid (PL) levels were assessed by using enzymatic kits from Wako Diagnostics (Richmond, VA). Fractions 10 to 15 were determined as VLDL by high content of TG, 16 to 22 as LDL by the lower protein peak, 25 to 36 as HDL by higher PL and TC content, and 37 to 45 as albumin. HDL fractions were evenly clustered into 3 pools according to particle size, fractions 25 to 28 as large-HDL particles (L-HDL), 29 to 32 as medium-HDL (M-HDL), and 33 to 36 as small-HDL (S-HDL). Total protein concentration in small, medium and large (S/M/L)-HDL fractions were assessed by BCA assay. Total PL, cholesterol, and TG in S/L/M-HDL fractions were normalized by protein content in each fraction.

Zhang Y., Gordon S.M., Xi H., Choi S., Paz M.A., Sun R., Yang W., Saredy J., Khan M., Remaley A.T., Wang J.F., Yang X, & Wang H. (2019). HDL subclass proteomic analysis and functional implication of protein dynamic change during HDL maturation. Redox Biology, 24, 101222.

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Pemafibrate Enhances FGF21 Signaling

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Pemafibrate was kindly provided by Kowa Co. Ltd (Nagoya, Japan). Mouse CD31 antibody was purchased from BD Pharmingen (San Jose, CA)(550274). Antibodies of phosphorylated eNOS (Ser-1177)(9571), eNOS (32027) and Tubulin (2144) were purchased from Cell Signaling Technology (Beverly, MA). NG-nitro-l-arginine methyl ester (L-NAME) was purchased from Sigma (St. Louis, MO) (N5751). GW6471 was purchased from Cayman Chemical (11697). Recombinant human FGF21 protein was purchased from R&D system (2539-FG-025). Plasma FGF21 levels were measured by ELISA kit (R & D system)(MF2100) [15 (link)]. Plasma adiponectin levels were determined by ELISA kit (Otsuka Pharmaceutical Co. Ltd.)(410713). Adenoviral vectors expressing mouse full-length FGF21 (Ad-FGF21) were constructed under the control of the CMV promoter [16 (link), 17 (link)]. Adenoviral vectors expressing β-galactosidase (Ad-βgal) were used as controls [18 (link)]. Lipid profiles and plasma glucose were analyzed by enzymatic kits (Wako Pure Chemical Industries, Ltd) (total cholesterol, 439–17501) (triglyceride, 290–63701) (glucose, 439–90901).

Kawanishi H., Ohashi K., Ogawa H., Otaka N., Takikawa T., Fang L., Ozaki Y., Takefuji M., Murohara T, & Ouchi N. (2020). A novel selective PPARα modulator, pemafibrate promotes ischemia-induced revascularization through the eNOS-dependent mechanisms. PLoS ONE, 15(6), e0235362.

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Fasting Mouse Serum Metabolite Analysis

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Blood samples were collected from mice in the fasting (14 hours) state. Serum glucose, total cholesterol and triglyceride concentrations were measured with enzymatic kits (Wako Pure Chemical Industries, Ltd.).

Yoshida S., Fuster J.J, & Walsh K. (2014). Adiponectin attenuates abdominal aortic aneurysm formation in hyperlipidemic mice. Atherosclerosis, 235(2), 339-346.

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Lipid Profile Analysis in Mice

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Lipid levels were analyzed in plasma and liver samples. Blood was drawn via the tail vein from mice fasted for 5 h. Total plasma cholesterol and TG levels were determined using commercially available kits (Sekisui Diagnostics). Cholesterol in VLDL fractions was measured with the Amplex Red cholesterol assay kit (Thermo Fisher Scientific). Plasma NEFA and 3-hydroxybutyrate levels were determined using enzymatic kits (Wako Chemicals). Liver samples were homogenized using a hypotonic extraction buffer [250 mM sucrose, 5 mM Tris, 1 mM EDTA, and proteinase inhibitor (cOmplete; Sigma)] and total cholesterol and TG were determined in the supernatant after centrifugation as described above.

Ramms B., Patel S., Nora C., Pessentheiner A.R., Chang M.W., Green C.R., Golden G.J., Secrest P., Krauss R.M., Metallo C.M., Benner C., Alexander V.J., Witztum J.L., Tsimikas S., Esko J.D, & Gordts P.L. (2019). ApoC-III ASO promotes tissue LPL activity in the absence of apoE-mediated TRL clearance. Journal of Lipid Research, 60(8), 1379-1395.

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Serum Lipid and Inflammation Analysis

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Serum total cholesterol, high-density lipoprotein cholesterol, non-esterified fatty acids (NEFAs) and triglycerides were measured by enzymatic kits purchased from Wako Diagnostics (Richmond, VA, Canada). Plasma glucose was measured by an enzymatic assay acquired from Pointe Scientific, Inc (Canton, MI, USA). In addition, serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured by spectrophotometric methods (Pointe Scientific, Inc; Canton, MI, USA). Following kit instructions, indirect sandwich ELISAs purchased from R&D Systems (Minneapolis, MN, USA) were used to measure the inflammatory markers, interleukin-1-beta (IL-1β), MCP-1, and serum amyloid A (SAA).

Millar C.L., Norris G.H., Vitols A., Garcia C., Seibel S., Anto L, & Blesso C.N. (2019). Dietary Egg Sphingomyelin Prevents Aortic Root Plaque Accumulation in Apolipoprotein-E Knockout Mice. Nutrients, 11(5), 1124.

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Liver Lipid Analysis and Histology

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Liver lipids were extracted using methods previously described (19 (link)) with a modified Folch method. Following extraction and solubilization in 1% Triton X-100, total cholesterol, free cholesterol, and TGs were measured using enzymatic kits (Wako Diagnostics, Richmond, VA). Cholesteryl esters were also calculated as (total cholesterol − free cholesterol) × 1.67.
For liver histology, fresh tissue sections were placed in cassettes and fixed in 10% neutral-buffered formalin. Samples were processed and stained with hematoxylin and eosin at the Connecticut Veterinary Medical Diagnostic Laboratory in Storrs, CT, as previously reported (19 (link)). Slides were imaged and used to present representative images of the tissue histology in each group.

Millar C.L., Anto L., Garcia C., Kim M.B., Jain A., Provatas A.A., Clark R.B., Lee J.Y., Nichols F.C, & Blesso C.N. (2022). Gut microbiome-derived glycine lipids are diet-dependent modulators of hepatic injury and atherosclerosis. Journal of Lipid Research, 63(4), 100192.

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Lipid Quantification Protocol

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Plasma total cholesterol, free fatty acid, triglyceride and HDL concentrations were measured using enzymatic kits (Wako Chemicals, Richmond VA.).

Ji A., Trumbauer A.C., Noffsinger V.P., Jeon H., Patrick A.C., De Beer F.C., Webb N.R., Tannock L.R, & Shridas P. (2022). Serum Amyloid A is not obligatory for high-fat, high-sucrose, cholesterol-fed diet-induced obesity and its metabolic and inflammatory complications. PLoS ONE, 17(4), e0266688.

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