蛋白酶K(Proteinase K) 蛋白酶k水溶液

蛋白酶K(Proteinase K) 蛋白酶k水溶液

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产品简介

 

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29 kDa,可切割脂肪族氨基酸和芳香族氨基酸的羧基端肽键。Proteinase K 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 50-100 µg/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。

 

产品信息

 

货号

10401JP60/ 10401JP80 /10401JP95/ 10401JP98

规格

100 mg /1 g/10 g/100 g

 

组分性质

 

1)本品来源于林伯氏白色念球菌(Tritirachium album);

2)比活性值(Specific activity):≥30 U/mg 蛋白;

3)不含 RNase 和 DNase;

4)在尿素和 SDS 中活性稳定存在;

5)在较广的 pH 范围内(pH 4.5 – 12.5)均有活性,最佳 pH 范围为 pH 7.0 12.0

6)反应条件:反应缓冲液中 37℃ – 65℃温育 1 h。推荐反应温度选择 65℃;

7)活力单位定义:在规定条件下,每分钟水解酪蛋白生成 1 μmol L-酪氨酸所需的酶量;

8)失活方法:反应体系中加入 PMSF 或 DFIP 抑制剂即可失活。

 

储存条件

 

-25~-15 ℃干燥储存,有效期 3 

 

注意事项

 

1)为了您的安全和健康,请穿实验服并戴一次性手套操作。

2)启用后如需反复使用,需置于-20℃~-80℃冰箱冷冻或者配制成溶液后过滤除菌分装保存。

3)本产品仅作科研用途!

 

使用说明

 

1. 20-40 mg/mL Proteinase K 储存液的配制

1)将干粉溶解于稀释缓冲液中,配制成 40-80 mg/mL 原液;

【注】:稀释缓冲液配方:20 mM Tris-HCl (pH 7.4), 1 mM CaCl2

2)使用低蛋白吸附的 0.22 µm 滤器过滤除菌;

3)加入等体积无菌甘油,配制成含 50%甘油的 Proteinase K 储存液。

【注】:Proteinase K 储存液 4℃保存2年。若需反复使用,需分装保存。

2. 按照核酸提取说明书添加指定量Proteinase K储存液即可。

Ver.CN20231201

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)

产品简介

 

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29 kDa,可切割脂肪族氨基酸和芳香族氨基酸的羧基端肽键。Proteinase K 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 50-100 µg/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。

 

产品信息

 

货号

10401JP60/ 10401JP80 /10401JP95/ 10401JP98

规格

100 mg /1 g/10 g/100 g

 

组分性质

 

1)本品来源于林伯氏白色念球菌(Tritirachium album);

2)比活性值(Specific activity):≥30 U/mg 蛋白;

3)不含 RNase 和 DNase;

4)在尿素和 SDS 中活性稳定存在;

5)在较广的 pH 范围内(pH 4.5 – 12.5)均有活性,最佳 pH 范围为 pH 7.0 12.0

6)反应条件:反应缓冲液中 37℃ – 65℃温育 1 h。推荐反应温度选择 65℃;

7)活力单位定义:在规定条件下,每分钟水解酪蛋白生成 1 μmol L-酪氨酸所需的酶量;

8)失活方法:反应体系中加入 PMSF 或 DFIP 抑制剂即可失活。

 

储存条件

 

-25~-15 ℃干燥储存,有效期 3 

 

注意事项

 

1)为了您的安全和健康,请穿实验服并戴一次性手套操作。

2)启用后如需反复使用,需置于-20℃~-80℃冰箱冷冻或者配制成溶液后过滤除菌分装保存。

3)本产品仅作科研用途!

 

使用说明

 

1. 20-40 mg/mL Proteinase K 储存液的配制

1)将干粉溶解于稀释缓冲液中,配制成 40-80 mg/mL 原液;

【注】:稀释缓冲液配方:20 mM Tris-HCl (pH 7.4), 1 mM CaCl2

2)使用低蛋白吸附的 0.22 µm 滤器过滤除菌;

3)加入等体积无菌甘油,配制成含 50%甘油的 Proteinase K 储存液。

【注】:Proteinase K 储存液 4℃保存2年。若需反复使用,需分装保存。

2. 按照核酸提取说明书添加指定量Proteinase K储存液即可。

Ver.CN20231201

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)