重组胰蛋白酶是由重组大肠杆菌表达生产,氨基酸序列与猪胰腺来源的阳离子型胰蛋白酶完全一致,无糜蛋白酶等杂酶污染,不含有且不需要TPCK抑制其他酶活性,活性高,酶切特异性高。胰蛋白酶容易自切产生自身自切片段影响质谱分析结果,甲基化修饰和突变保证其不自切,从而也去除了因自切而产生的假糜蛋白酶的活性。
推荐使用方法
建议使用50mMHAc或1mM HCl溶解或稀释。使用时,稀释在50mM NH4HCO3 或者ph7.0-8.0的缓冲溶液中。在酶切缓冲液中建议使用1mM CaCl2,重组胰蛋白酶:目的蛋白=1:20-1:100,最适PH为7.0-8.0。
产品优势
序列分析纯蛋白酶:特异性高,稳定性好。
无动物源性:重组生产,无外源性的病毒污染,生产过程不使用任何动物源原料。
质量稳定:批量生产,可保证稳定连续的批次生产;产 品批次间无差异,质量稳定。
纯度高:比活高;宿主蛋白残留小于生物制品限度要求。
冻干粉:易于储存和运输。
产品用途
重组胰蛋白酶具有与动物源性胰蛋白酶相同的酶学性质。可替代胰蛋白酶使用,且有很多优势,如无杂酶活性,高稳定性,不自切,高活性;在使用中,无非特异性酶切片段及自切片段出现。用于特异性的蛋白质酶解、蛋白测序、制作肽谱图、蛋白质组学研究、Z-D胶上的肽段的胰酶消化等。
Recombinant trypsin is expressed and produced by recombinant Escherichia coli. Its amino acid sequence is identical to that of cationic trypsin derived from porcine pancreas. It is free of contaminating enzymes such as chymotrypsin; TPCK is neither contained nor required to inhibit interfering enzymatic activities. It features high activity and stringent digestion specificity.
Conventional trypsin is prone to autolysis, generating autolytic fragments that interfere with mass spectrometry results. Methylation modification and site-directed mutations eliminate its autolytic capacity, and consequently abolish pseudo-chymotrypsin activity arising from autodegradation.
Recommended Protocol
Dissolve or dilute the product with 50 mM HAc or 1 mM HCl. For digestion, further dilute it in 50 mM NH₄HCO₃ or buffer solutions at pH 7.0–8.0.
Add 1 mM CaCl₂ to the digestion buffer as recommended. The mass ratio of recombinant trypsin to target protein ranges from 1:20 to 1:100, with an optimal pH of 7.0–8.0.
Product Advantages
1. Sequence-grade pure protease: Excellent substrate specificity and superior stability.
2. Animal-origin-free: Manufactured via recombinant expression without any animal-derived raw materials, eliminating risks of exogenous viral contamination.
3. Consistent quality: Mass production enables stable continuous batch manufacturing with negligible inter-batch variability.
4. High purity & specific activity: Residual host cell protein levels are below the limits specified for biological products.
5. Lyophilized powder format: Convenient for storage and transportation.
Applications
Recombinant trypsin shares identical enzymatic properties with animal-derived trypsin and serves as a superior alternative thanks to the absence of contaminating enzymatic activities, enhanced stability, resistance to autolysis, and high catalytic activity. No non-specific cleavage fragments or autolytic byproducts are generated during digestion.
It is widely applied in site-specific protein digestion, protein sequencing, peptide mapping, proteomics research, and in-gel tryptic digestion of proteins separated by SDS-PAGE.