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重组嗜酸细菌Cas12b蛋白
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重组嗜酸细菌Cas12b蛋白

重组嗜酸细菌Cas12b蛋白,Cas12b, Recombinant, A. acidiphilus,非常适合开展酶动力学,抑制剂筛选和选择性分析实验类型。更多视频请关注视频号【艾维缔】。哔哩哔哩【IVDSHOW】。抖音【军哥聊表观】。快手:【表观盒子®】;小红书:【EpiBox®】
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AapCas12属于V型CRISPR效应子CRISPR-Cas12b/C2c1,具有很高的热稳定性,可广泛应用。在大肠杆菌中表达的与Alicyclobacillus acidiphilus Cas12b(V型CRISPR相关蛋白Cas12b)相对应的全长重组蛋白。

Molecular Weight

~128kD

Assay Conditions

Varying amounts of AapCas12b activity was measured using a CRISPR-based fluorescent reporter assay for optimal results. Target DNA cutting and indiscriminate single stranded DNA collateral cleavage was activated using RNA-guided DNA Binding to Cas12. Emission of fluorescent signal is due to the degradation of ssDNA reporters upon cleavage.

Active Cas12 was thawed on ice while 1X Endonuclease Buffer containing 10 mM Tris-HCl, pH 8.0, 50 mM NaCl, 10 mM MgCl2, and 0.1 mg/ml BSA, guide RNA (custom designed crRNA), ds DNA activator (complementary sequence to crRNA and a PAM sequence specific for Cas enzyme) and FQ-ssDNA substrate (labeled with fluorophore and a quencher) were equilibrated to room temperature. Next three working solutions of Active Cas12 (4X final concentration) guide RNA (4X final concentration) and activator/reporter mix containing ds DNA activator and ssDNA reporter (2X final concentration), were prepared using 1X Endonuclease Buffer. 10 μl of 4X active Cas12 and 10 μl of 4x guide RNA were then preincubated in half the area of a solid black 96-well plate for 10 minutes at room temperature. After preincubation, 20 μl of 2X activator/reporter mix was added to plate and placed on shaking incubator for 1 min. The plate was then sealed and incubated at 37°C for 10-30 minutes. Plate was then equilibrated to room temperature, plate sealer removed and fluorescence read on a microplate reader. Negative control was measured by replacing enzyme working solution with equal volume of assay buffer.

Applications

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Storage and Stability

Aliquot to avoid repeated freezing and thawing and store at -70°C. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Source

Recombinant, E. coli

Purity

≥75%

Concentration

As Reported

Form

Supplied as a liquid in 50mM sodium phosphate, pH 7.5, 300mM sodium chloride, 1mM DTT, 10% glycerol

Grade

Purified

Accession Number

WP_067623834

Molecular Weight

128

产品组分

内容

型号

规格

储存温度

重组嗜酸细菌Cas12b蛋白 045812 20ug -70°C

操作手册

1 1

常温

文件资源

文件下载 文件内容 资源说明
045812-重组嗜酸细菌Cas12b蛋白 操作手册
045812-重组嗜酸细菌Cas12b蛋白相关单页 宣传单页

 

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注意事项

保存建议 厂家推荐蓝冰运输。当您收到产品后,按照说明书建议保存于-20°C。

 

FAQ

1. Ocampo Lyra J.S.P., Madi K., Maeda C.T., Savino W. (1993) Thymic extracellular matrix in human malnutrition. J. Pathol. 1993, 171, 231-236.

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