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Tac assay kit

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The TAC Assay Kit is a laboratory tool used to quantitatively measure the total antioxidant capacity (TAC) of biological samples. It provides a simple and reliable method to assess the overall antioxidant status of a sample.

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Lab products found in correlation

Mineral oil, by Merck Group (1 mentions) Uric acid, by Merck Group (1 mentions) Whatman grade 1 qualitative filter paper, by GE Healthcare (1 mentions) Gastight 1750, by Hamilton Company (1 mentions) Enspire 2300, by PerkinElmer (1 mentions) 11 pico plus elite, by Harvard Apparatus (1 mentions) Duoset elisa, by R&D Systems (1 mentions) Human igf 1 quantikine elisa kit, by R&D Systems (1 mentions) Oxiselect nitrotyrosine elisa kit, by Cell Biolabs (1 mentions) Gsh gssg glo assay kit, by Promega (1 mentions)

6 protocols using tac assay kit

1

Plasma Total Antioxidant Capacity Assay

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Plasma total antioxidant capacity (TAC) was determined using TAC Assay Kit (Cell Biolabs OxiSelect™, San Diego, California, USA). This assay is based on the reduction of copper (II) to copper (I) by uric acid. Upon reduction, the copper (I) ion further reacts with a coupling chromogenic reagent that produces a color that measured at 490 nm. Absorbance values were compared with a known uric acid standard curve. The assay processes were achieved following manufacturers assay techniques. TAC was expressed as mM.
Harisa G.I., Attia S.M., Zoheir K.M, & Alanazi F.K. (2016). Chitosan treatment abrogates hypercholesterolemia-induced erythrocyte’s arginase activation. Saudi Pharmaceutical Journal : SPJ, 25(1), 120-127.
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2

Colorimetric Total Antioxidant Capacity Assay

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The TAC assay kit adopted for TAC tests on cPADs was purchased from Cell Biolabs, Inc., USA. Food dye, a product of McCormick (USA), was diluted 20-fold with deionized water. A 60 mM uric acid (UA, Sigma-Aldrich, USA) stock solution was prepared freshly in 1 M NaOH and then serially diluted into 4, 2, 1, 0.5, 0.25 and 0.125 mM solutions with 18 MΩ DI water. Citrate concentrated solution (4% w/v, Sigma-Aldrich, USA) was mixed with whole blood at a volume ratio of 1:9 to prevent blood from coagulation. Mineral oil (Sigma-Aldrich, USA) was used as the carrier of the blood plug.
Whatman® Grade 1 Qualitative Filter Papers (GE Healthcare, UK) and Brand® Parafilm® M sealing film (Bemis Company, Inc., USA) were used to make cPADs. Craft punches are products of Fiskars Brands, Inc. (USA). A 10-cm long PTFE tubing (365-μm i.d., 800-μm o.d.) connected to a 500-μL syringe (Gastight 1750, Hamilton Company, USA) was taken for plasma separation. A syringe pump (11 Pico Plus Elite, Harvard Apparatus, USA) provided the driving force for the flow. A microplate reader (EnSpire 2300, PerkinElmer, USA) was used to measure TAC on a 96-well plate (BD Falcon, USA). cPADs images were captured by a scanner (HP C4480, USA) and analyzed with the ImageJ software package (NIH, USA).
Sun M, & Johnson M.A. (2015). Measurement of Total Antioxidant Capacity in Sub-μL Blood Samples Using Craft Paper-based Analytical Devices. RSC advances, 5(69), 55633-55639.
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3

Evaluating Antioxidant Capacity in ASC-CM

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Collected ASC-CM sample antioxidant activity was assessed with a Total Antioxidant Capacity (TAC) Assay kit (Cell Biolabs; Cat. #STA-360), per the manufacturer's instructions. The TAC kit evaluates antioxidant activity via the reduction of copper (II) to copper (I) and utilizes the naturally occurring antioxidant uric acid as a control standard. Thus, antioxidant activity was displayed as “mM equivalents” of uric acid. Control serum-free MSC media was used to assess baseline antioxidant activity of media without exposure to cells. Assays were performed with technical replicates and biological triplicates (n = 3).
Hodge J.G., Robinson J.L, & Mellott A.J. (2023). Tailoring the secretome composition of mesenchymal stem cells to augment specific functions of epidermal regeneration: an in vitro diabetic model. Frontiers in Medical Technology, 5, 1194314.
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4

Biomarker Assessment in Fasting Blood

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Fasting blood samples were collected from the jugular vein at baseline and monthly. Blood was collected in 0.1 mol/L citrate-containing EDTA. Fresh whole blood was submitted to Antech Diagnostics for CBC with differential and biochemistry measurements (Superchem w/CBC, SA020). Plasma IGF-1 levels were quantified by human IGF-I Quantikine ELISA Kit (R&D Systems, DG100B) and CRP levels by porcine C-reactive protein/CRP DuoSet ELISA (R&D Systems, DY2648). Quantification of plasma N-tyrosine and TAC assay were performed with OxiSelect Nitrotyrosine ELISA Kit (STA-305) and TAC assay kit (STA-360) (both from Cell Biolabs Inc).
Sukhanov S., Higashi Y., Yoshida T., Danchuk S., Alfortish M., Goodchild T., Scarborough A., Sharp T., Jenkins J.S., Garcia D., Ivey J., Tharp D.L., Schumacher J., Rozenbaum Z., Kolls J.K., Bowles D., Lefer D, & Delafontaine P. (2023). Insulin-like growth factor 1 reduces coronary atherosclerosis in pigs with familial hypercholesterolemia. JCI Insight, 8(4), e165713.
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5

Quantifying Antioxidant Levels in Prostate Cancer Cells

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The amounts of total glutathione in PC-3 and DU145 cells were estimated using the GSH/GSSG-Glo™ Assay Kit (Promega), according the manufacturer's instructions. Such a luminescence-based system allows the detection and quantification of total GSH and GSSG as well as the GSH/GSSG ratios in cultured cells by comparison with a GSH standard curve.
The Total Antioxidant Capacity (TAC) of PC-3 and DU145 cells was measured by a TAC Assay Kit (Cell Biolabs Inc., San Diego, CA), according to the manufacturer's protocol. Briefly, 24 h after seeding the cells were collected, washed and lysed by sonication in PBS. Cell lysates were then diluted with a reaction reagent and, upon addition of Cu 2+ solution, the reaction was stopped and read with a spectrophotometric microplate reader at 490 nm. Antioxidant capacity, expressed as copper reducing equivalent (CRE), was determined by using a standard curve generated by using fixed concentrations of uric acid.
Beretta G.L., Folini M., Cavalieri F., Yan Y., Fresch E., Kaliappan S., Hasenöhrl C., Richardson J.J., Tinelli S., Fery A., Caruso F, & Zaffaroni N. (2015). Unravelling "off-target" effects of redox-active polymers and polymer multilayered capsules in prostate cancer cells. Nanoscale, 7(14).
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6

Measuring Antioxidant Capacity in Black Rice

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The Total Antioxidant Capacity (TAC) Assay Kit (Cell Biolabs OxiSelect™, San Diego, CA, USA) was used to measure the antioxidant capacity of ANT from black rice. The TAC Assay is based on the reduction of copper (II) to copper (I) by antioxidants such as uric acid. Briefly, RDFs were treated with 5–50 μg/mL ANT and stimulated with 0.3 mM H2O2 for 24 h. The supernatant was dispensed in a 96-well microtiter plate, after which 180 μL of 1× reaction buffer was added to each well and mixed. An initial reading at 490 nm was taken for each sample. Then, 50 μL of 1× copper ion reagent was added and incubated for 5 min on an orbital shaker. Next, 50 μL of stop solution was added to terminate the reaction, and the plate was subjected to additional measurement of absorption at 490 nm using a spectrophotometer (Epoch, BioTek, Winooski, VT, USA) with the Gen 5 data analysis software interface. All determinations were performed in triplicate, and results were averaged. The copper ion reduction activity of each sample with H2O2 was then calculated as the percent increase in ion reduction.
Palungwachira P., Tancharoen S., Phruksaniyom C., Klungsaeng S., Srichan R., Kikuchi K, & Nararatwanchai T. (2019). Antioxidant and Anti-Inflammatory Properties of Anthocyanins Extracted from Oryza sativa L. in Primary Dermal Fibroblasts. Oxidative Medicine and Cellular Longevity, 2019, 2089817.
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