Proteinase K（蛋白酶 K）是一种切割活性较广的丝氨酸蛋白酶，其相对分子量约为29kDa，可切割脂肪族氨基酸和芳香族氨基酸的羧基端肽键。ProteinaseK 应用广泛，可用于制备脉冲电泳的染色体 DNA，蛋白质印迹以及去除 DNA 和 RNA制备中的核酸酶等实验，而变性剂如 SDS（1%）可提高其活性，常用工作浓度为 50-100µg/mL，根据所用缓冲液是否含有SDS、尿素、pH、温度等因素确定具体的工作浓度。
- 源于基因重组酵母菌株。
- 不含 RNase 和 DNase。
- 在尿素和 SDS 中活性稳定存在。
- 在较广的 pH 范围内（pH 4.0 – 12.0）均有活性，最佳 pH 范围为 pH 7.5 - 9.0。
冰袋运输。2~8℃干燥保存，有效期 3 年。更长期保存，可置于-25~-15℃冷冻保存。
Q：蛋白酶 K 主要有哪些应用？
A：原位杂交样本制备；核酸纯化等实验蛋白质的切割清除。
[1] Li W, Qiu S, Chen J, et al. Chimeric Antigen Receptor Designed to Prevent Ubiquitination and Downregulation Showed Durable Antitumor Efficacy. Immunity. 2020;53(2):456-470.e6. doi:10.1016/j.immuni.2020.07.011(IF:22.553)
[2] Zhang Z, Mou Z, Xu C, et al. Autophagy-associated circular RNA hsa_circ_0007813 modulates human bladder cancer progression via hsa-miR-361-3p/IGF2R regulation. Cell Death Dis. 2021;12(8):778. Published 2021 Aug 7. doi:10.1038/s41419-021-04053-4(IF:8.469)
[3] Ni H, Qin H, Sun C, et al. MiR-375 reduces the stemness of gastric cancer cells through triggering ferroptosis. Stem Cell Res Ther. 2021;12(1):325. Published 2021 Jun 5. doi:10.1186/s13287-021-02394-7(IF:6.832)
[4] Chen S, Jia Z, Cai M, et al. SP1-Mediated Upregulation of Long Noncoding RNA ZFAS1 Involved in Non-syndromic Cleft Lip and Palate via Inactivating WNT/β-Catenin Signaling Pathway. Front Cell Dev Biol. 2021;9:662780. Published 2021 Jun 29. doi:10.3389/fcell.2021.662780(IF:6.684)
[5] Ma J, Chen S, Hao L, et al. Hypermethylation-mediated down-regulation of lncRNA TBX5-AS1:2 in Tetralogy of Fallot inhibits cell proliferation by reducing TBX5 expression. J Cell Mol Med. 2020;24(11):6472-6484. doi:10.1111/jcmm.15298(IF:4.486)
[6] Shi L, Gong Y, Zhuo L, Wang S, Chen S, Ke B. Methyltransferase-like 3 upregulation is involved in the chemoresistance of non-small cell lung cancer. Ann Transl Med. 2022;10(3):139. doi:10.21037/atm-21-6608(IF:3.932)
[7] Ma J, Chen S, Hao L, et al. Long non-coding RNA SAP30-2:1 is downregulated in congenital heart disease and regulates cell proliferation by targeting HAND2. Front Med. 2021;15(1):91-100. doi:10.1007/s11684-020-0778-5(IF:3.297)
[8] Huang Q, Xie Y, Yang Z, et al. The cytoplasmic loops of AgrC contribute to the quorum-sensing activity of Staphylococcus aureus. J Microbiol. 2021;59(1):92-100. doi:10.1007/s12275-021-0274-x(IF:2.845)
[9] Guo T, Xing Y, Chen Z, et al. Core-binding factor beta is required for osteoblast differentiation during fibula fracture healing. J Orthop Surg Res. 2021;16(1):313. Published 2021 May 14. doi:10.1186/s13018-021-02410-9(IF:2.359)
[10] Zhang W, Zhang T, Yan Y, et al. Exosomal miR-22-3p Derived from Chronic Rhinosinusitis with Nasal Polyps Regulates Vascular Permeability by Targeting VE-Cadherin. Biomed Res Int. 2020;2020:1237678. Published 2020 Nov 12. doi:10.1155/2020/1237678(IF:2.276)
[11] Cheng L, Yi X, Shi Y, et al. Abnormal lipid metabolism induced apoptosis of spermatogenic cells by increasing testicular HSP60 protein expression. Andrologia. 2020;52(11):e13781. doi:10.1111/and.13781(IF:1.951)
[12] Zhang T, Zhou Y, You B, et al. miR-30a-5p Inhibits Epithelial-to-Mesenchymal Transition by Targeting CDK6 in Nasal Polyps. Am J Rhinol Allergy. 2021;35(2):152-163. doi:10.1177/1945892420939814(IF:1.943)
