Nitrotetrazolium Blue chloride

Nitrotetrazolium Blue chloride (NBT) is a dehydrogenase substrate that is used in western blots and immunohistochemical staining with the alkaline phosphatase (ALP) substrate 5-bromo-4-chloro-3-indolyl phosphate (BCIP) to produce a purple-blue precipitate^[1]. Nitrotetrazolium Blue chloride can visualize the localization of target mRNAs at cellular resolution in embryos or tissues ^[2]. Nitrotetrazolium Blue chloride can react with reactive oxygen species (ROS) and superoxide anions (O₂^-) in cells or tissues to produce dark blue insoluble formazan compounds ^{[3, 4]}. Nitrotetrazolium Blue chloride can stain intracellular ALP and succinate dehydrogenase (SDH) ^{[5, 6]}.

References:
[1]Santangelo K S, Schaefer D M W, Leavell S E, et al. Special Staining Techniques: Application and Quality Assurance[J]. Veterinary Cytology, 2020: 47-72.
[2]Narayanan R, Oates A C. Detection of mRNA by whole mount in situ hybridization and DNA extraction for genotyping of zebrafish embryos[J]. Bio-protocol, 2019, 9(6): e3193-e3193.
[3]Lamacchia M, Dyrka W, Breton A, et al. Overlapping Podospora anserina transcriptional responses to bacterial and fungal non self indicate a multilayered innate immune response[J]. Frontiers in microbiology, 2016, 7: 471.
[4]Kumar D, Yusuf M A, Singh P, et al. Histochemical detection of superoxide and H2O2 accumulation in Brassica juncea seedlings[J]. Bio-protocol, 2014, 4(8): e1108-e1108.
[5]Yang C S, Gu Z T, Ding R, et al. Retracted: Long non‐coding RNA MEG3 silencing and microRNA‐214 restoration elevate osteoprotegerin expression to ameliorate osteoporosis by limiting TXNIP[J]. Journal of cellular and molecular medicine, 2021, 25(4): 2025-2039.
[6]Wang G, Biswas A K, Ma W, et al. Metastatic cancers promote cachexia through ZIP14 upregulation in skeletal muscle[J]. Nature medicine, 2018, 24(6): 770-781.

Nitrotetrazolium Blue chloride（NBT）是一种脱氢酶底物，与碱性磷酸酶（ALP）底物5-溴-4-氯-3-吲哚基磷酸酯（BCIP）一起用于蛋白质印迹和免疫组化染色，产生紫蓝色沉淀^[1]。Nitrotetrazolium Blue chloride可以在胚胎或组织中以细胞分辨率可视化靶mRNA的定位^[2]。Nitrotetrazolium Blue chloride可以在细胞或组织中与活性氧（ROS）和超氧阴离子（O₂^-）反应生成深蓝色不溶性甲臜化合物^{[3, 4]}。Nitrotetrazolium Blue chloride可以对细胞内ALP和琥珀酸脱氢酶（SDH）染色^{[5, 6]}。