Abcam | 活性氧物質 (reactive oxygen species, ROS) 與氧化壓力 (oxidative stress) 檢測

Abcam | 活性氧物質 (reactive oxygen species, ROS) 與氧化壓力 (oxidative stress) 檢測

2024.04.29

活性氧與氧化壓力檢測介紹

活性氧(ROS)是含有氧的活性化學物質。這些包括過氧化物、超氧化物、羥基自由基、單線態氧和α-氧。

由於 ROS 的瞬態性質,儘管使用基於螢光染料的檢測(例如 DCFDA)可以輕鬆在活細胞中測量它們,但在組織和生物流體樣本中測量它們可能更困難。

DCFDA 檢測方案是基於 DCFDA/H2DCFDA/DCFH-DA/DCFH 擴散到細胞。進入細胞後,它被細胞酯酶去乙醯基成非螢光化合物,並被ROS氧化成2′,7′-二氯螢光素(DCF)。 DCF 具有強螢光性,可透過激發/發射波長為 485/535 nm 的螢光光譜進行檢測。

Cat no. Product name Sample type Platform
ab113851 DCFDA / H2DCFDA – Cellular ROS Assay Kit Adherent cells, Suspension cells Microplate reader, Flow cytometer, Fluorescence microscope
ab139477 Cellular Superoxide Detection Assay Kit Adherent cells, Suspension cells Flow cytometer, Fluorescence microscope
ab139476 ROS/Superoxide Detection Assay Kit (Cell-based) Adherent cells, Suspension cells Flow cytometer, Fluorescence microscope
ab186027 Cellular ROS Assay Kit (Red) Adherent cells, Suspension cells Microplate reader, Fluorescence microscope
ab186028 Cellular ROS Assay Kit (Orange) Adherent cells, Suspension cells Microplate reader, Flow cytometer, Fluorescence microscope
ab186029 Cellular ROS Assay Kit (Deep Red) Adherent cells, Suspension cells Microplate reader, Flow cytometer, Fluorescence microscope
ab139473 Cellular ROS/RNS Assay Kit Adherent cells, Suspension cells Fluorescence microscope
ab219931 Mitochondrial Hydroxyl Radical Detection Assay Kit Adherent cells, Suspension cells Microplate reader, Fluorescence microscope
ab219943 Mitochondrial Superoxide Assay Kit (Fluorometric) Adherent cells, Suspension cells Microplate reader, Fluorescence microscope
ab138874 Hydrogen Peroxide Assay Kit (Cell-based) Adherent cells, Suspension cells Fluorescence microscope
ab138886 Hydrogen Peroxide Assay Kit (Fluorometric – Near Infrared) Adherent cells, Suspension cells Microplate reader
ab102500 Hydrogen Peroxide Assay Kit Cell culture media, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine Microplate reader

DNA/RNA 損傷、脂質過氧化和蛋白質氧化/硝化

氧化壓力(oxidative stress)可以透過測量 DNA/RNA 損傷、脂質過氧化和蛋白質氧化/硝化的水平來間接測量,而不是直接測量 ROS。這些氧化壓力標記物比 ROS 具有更持久的特性。

  • DNA/RNA損傷

有幾種類型的 DNA/RNA 損傷可以作為氧化壓力的標記進行測量。 8-羥基脫氧鳥苷 (8-OHdG) 可能是測量氧化壓力最常用的 DNA 損傷標記物。彗星試驗、脫嘌呤/脫嘧啶位點試驗和醛誘導損傷試驗可以間接測量與氧化壓力相關的DNA損傷。

  • 脂質過氧化

丙二醛(MDA)是最常用的氧化壓力脂質標記。 MDA 透過多元不飽和脂肪酸的過氧化形成,通常使用 TBARS 測定進行定量。然而,其他醛類也在 TBARS 測定中產生訊號,因此該測定對於 MDA 並不完全具有特異性。儘管如此,TBARS 測定通常比使用 HPLC 測量 MDA 更方便。您也可以執行競爭性 ELISA 測定來測量 MDA。

其他脂質過氧化標記包括 4-HNA、8-異前列腺素、脂質氫過氧化物和氧化 LDL。

  • 蛋白質氧化/硝化

蛋白質引起的氧化損傷可以以蛋白質羰基化和蛋白質硝化(3-硝基酪胺酸)的形式發生。 ROS也可引起晚期糖化終產物(AGE)和高級氧化蛋白產物(AOPP)的形成。所有這些標記物都可以使用標準測定來測量。

Cat no. Product name Sample type Detection method
ab201734 8-hydroxy 2 deoxyguanosine ELISA Kit Cell Lysate, Plasma, Saliva, Serum, Tissue Extracts, Urine Colorimetric
ab133085 Lipid Hydroperoxide (LPO) Assay Kit Cell culture extracts, Plasma, Tissue Extracts Colorimetric
ab118970 Lipid Peroxidation (MDA) Assay Kit Cell culture extracts, Plasma, Tissue Extracts, Urine Colorimetric, Fluorometric
ab126287 Protein Carbonyl Content Assay Kit Cell Lysate Colorimetric
ab211154 DNA Damage Assay Kit (AP sites) DNA Colorimetric
ab178020 Protein Carbonyl Assay Kit (Western Blot) Adherent cells, Suspension cells, Tissue Homogenate Chemiluminescent

抗氧化劑 (Antioxidants)

抗氧化酶和其他還原分子可中和引起氧化損傷的活性氧。有三種類型的抗氧化劑用作氧化壓力標記物:小分子、酵素和蛋白質(例如白蛋白)。

有許多測定樣品總抗氧化能力的方法,包括測量小分子和蛋白質抗氧化能力的方法。 最常見的總抗氧化能力測定之一是 Trolox 當量抗氧化能力測定 (TEAC)。氧自由基抗氧化能力測定(ORAC)是另一種常見的氧化壓力分析方法,可透過減少ROS產生的螢光來測量抗氧化劑的能力。

抗氧化活性也可以透過測量特定分析物的量來確定。例如,這可以透過查看 GSH 和 GSSG 的相對量來完成。穀胱甘肽 (GSH) 被認為是內源性抗氧化劑中最豐富的分子,並形成氧化形式 GSSG。它被穀胱甘肽還原酶回收。

或者,可以根據氧化壓力水平來測量抗氧化酶(例如 GST 和超氧化物歧化酶)的活性水平。

Cat no. Product name Sample type Detection method
ab65329 Total Antioxidant Capacity Assay Kit Cell culture media, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine Colorimetric
ab234626 FRAP Assay Kit (Ferric Reducing Antioxidant Power Assay) Other biological fluids, Plasma, Serum Colorimetric
ab65656 Ascorbic Acid Assay Kit (Biological Samples) Cell culture extracts, Cell culture media, Cell culture supernatant, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine Colorimetric
ab65348 NAD/NADH Assay Kit Cell Lysate, Serum, Tissue Lysate, Urine Colorimetric
ab176723 NAD/NADH Assay Kit Cell Lysate, Tissue Lysate Fluorometric
ab65349 NADP/NADPH Assay Kit Cell Lysate, Tissue Extracts Colorimetric
ab176724 NADP/NADPH Assay Kit Cell culture extracts, Tissue Extracts Fluorometric
ab138881 GSH/GSSG Ratio Detection Assay Kit (Green) Cell Lysate, Plasma, Serum, Tissue Extracts, Urine Fluorometric
ab205811 GSH/GSSG Ratio Detection Assay Kit II (Green) Cell Lysate, Plasma, Tissue Extracts, Urine Fluorometric
ab112158 Free Thiol Assay Kit Cell culture extracts, Plasma, Urine Fluorometric
ab219272 Total Protein Thiol Assay Kit Purified protein Colorimetric
ab112132 Intracellular glutathione (GSH) Detection Assay Kit Adherent cells, Suspension cells Fluorometric
ab65325 GST Activity Assay Kit Cell Lysate, Other biological fluids, Plasma, Purified protein, Saliva, Serum, Tissue Lysate, Urine Fluorometric
ab65326 GST Assay Kit Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine Colorimetric
ab65354 Superoxide Dismutase Activity Assay Kit Cell culture media, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine Colorimetric
ab83461 Glutathione Reductase (GR) Assay Kit Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine Colorimetric
ab102522 Xanthine Oxidase Activity Assay Kit Cell culture extracts, Milk, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine Colorimetric, Fluorometric
ab102530 Glutathione Peroxidase Assay Kit Cell culture supernatant, Other biological fluids, Plasma, Platelets, Serum, Tissue Extracts, Urine Colorimetric
ab109712 Aconitase Activity Assay Kit Cell culture extracts, Tissue Colorimetric
ab118184 Catalase Activity Assay Kit (Immunocapture) Cell culture extracts, Tissue Extracts Colorimetric
ab83464 Catalase Activity Assay Kit Cell Lysate, Plasma, Serum, Tissue Lysate, Urine Colorimetric, Fluorometric
ab83463 Thioredoxin Reductase Assay Kit Cell culture extracts, Other biological fluids, Tissue Extracts, Urine Colorimetric
ab184867 NQO1 Activity Assay Kit Adherent cells, Cell culture extracts, Cell Lysate, Suspension cells, Tissue Extracts, Tissue Homogenate, Tissue Lysate Colorimetric
ab155895 Peroxidase Activity Assay Kit Cell culture supernatant, Other biological fluids, Plasma, Serum, Urine Colorimetric, Fluorometric
ab179843 Oxidative Stress Defense (Catalase, SOD1, TRX, smooth muscle Actin) Western Blot Cocktail Cell Lysate, Tissue Homogenate Chemiluminescent

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