关键区别 - 硝酸纤维素与PVDF
Western blotting是一种允许从蛋白质样品中检测和定量特定蛋白质的方法。该技术的可靠性取决于选择正确的膜以吸收凝胶中蛋白质的方法。有不同类型的微孔膜。硝酸纤维素和PVDF膜是研究人员首选的两个膜,因为它们的特殊属性比其他类型的膜。硝酸纤维素或PVDF之间的选择也是蛋白质印迹的另一个挑战。硝酸纤维素和PVDF均具有较高的蛋白质吸收能力。硝酸纤维素和PVDF膜之间的关键区别在于nitrocellulose membranes do not have the ability to strip off antibodies and reusing the membrane for antibody reprobing while PVDF membranes have stripping and reusing ability。
CONTENTS
1。概述和关键差异
2。What is Nitrocellulose
3。What is PVDF
4。Side by Side Comparison – Nitrocellulose vs PVDF
5。Summary
什么是硝酸纤维素?
硝酸纤维素为apolymermanufactured by treatingcellulosewith nitric acid and is used to make microporous membranes in molecular biology, especially for blotting techniques such as南部,北部和西部印迹。硝酸纤维素膜的孔径范围为3至20 µm。硝酸纤维素微孔膜有助于检测膜表面上发生的免疫化学反应。因此,硝酸纤维素膜经常用于固定蛋白质并检测蛋白质中的特定蛋白质。硝酸纤维素膜也可以固定糖蛋白和核酸。
硝酸纤维素膜在横向流程中首选测定due to several features. Nitrocellulose membranes absorb proteins at a high concentration. The solvent used to wet the membrane does not diminish the nitrocellulose membrane protein absorption. Nitrocellulose membranes can be easily cut to the desired gel size and transfer proteins from the gel to the membrane by electric or capillary transfer. Nitrocellulose allows faster flow of proteins through the membrane with high binding potential. Nitrocellulose shows an improved strength of handling. Another special attribute of nitrocellulose membrane is that it can be easily glued with non-solvent water resistant adhesives onto various plastic backings.
什么是PVDF?
Polyvinylidene difluoride (PVDF) is fluoropolymer produced by the polymerization of vinylidene difluoride and has a high immobilization capability of proteins. Therefore, microporous membranes made from PVDF are used in western blotting techniques to analyze specific proteins from the mixtures of proteins. PVDF membranes can also be used foramino acid分析和蛋白质测序。PVDF膜上硝酸纤维素膜上最重要的特征是,它可以轻松地从抗体中剥离并重复使用以进行后续抗体探针。
PVDFmembranes are thicker than nitrocellulose membranes; hence, more resistant to damages during reuse. PVDF membranes are highly疏水。Therefore, they must be soaked in甲醇或使用前异丙醇。
硝酸纤维素和PVDF有什么区别?
硝酸纤维素vs PVDF |
|
硝酸纤维素是由纤维素组成的聚合物。 | PVDFis a fluoropolymer produced by the polymerization of vinylidene difluoride. |
Membrane Pore Size | |
典型的孔径为0.1、0.2或0.45μ | 典型的孔径为0.1、0.2或0.45μm |
Protein Binding Capacity | |
硝酸纤维素has a protein binding capacity of 80 to 100 μg/cm2。 | PVDF的蛋白质结合能力为170至200μg/cm2。 |
灵敏度 | |
This has a low sensitivity compared to PVDF. | This has a high sensitivity. |
Detection of Lowly Expressed Proteins | |
由于硝酸纤维素膜的灵敏度低,因此不适合检测低表达的蛋白质。 | 由于其高灵敏度,这更适合于检测低表达的蛋白质。 |
背景噪音 | |
这具有较低的背景噪音 | 这具有更高的背景噪声。 |
与蛋白质的相互作用 | |
Protein molecules bind to nitrocellulose membranes through hydrophobic interactions. | 蛋白质通过疏水和偶极相互作用与PVDF膜结合。 |
膜的性质 | |
硝酸纤维素是脆弱的。但是,可以使用硝酸纤维素的版本,并且具有抗性。 | PVDF更耐用,具有更高的耐化学性。 |
Ability to Strip and Reuse | |
硝酸纤维素may have difficulty in stripping and reprobing without losing the signal. | PVDF非常适合重新操作和测序应用。 |
适应性 | |
硝酸纤维素is ideal for detecting low molecular weight proteins. | PVDF更适合检测较高的分子量蛋白。 |
其他用途 | |
硝酸纤维素可用于核酸分析和点/插槽印迹。 | PVDFcan be used for protein sequencing and solid phase assay systems. |
Cost | |
这比PVDF膜便宜。 | This is more expensive than nitrocellulose membranes. |
需要预润湿 | |
硝酸纤维素膜不需要用甲醇预设 | PVDF膜需要用甲醇预设。 |
Summary – Nitrocellulose vs PVDF
硝酸纤维素膜是商业上用于侧向流量测定法的第一个膜。它们具有吸收蛋白质的高容量。因此,硝酸纤维素膜用于蛋白质印迹。PVDF是蛋白质印迹中使用的另一种膜类型,它的蛋白质吸收能力也很高。两种类型都用于蛋白质印迹进行蛋白质分析。但是,PVDF膜具有更多特殊的属性,使其比硝酸纤维素膜更适合蛋白质印迹。但是,硝酸纤维素膜更适合于检测低分子加权蛋白,PVDF膜更适合于检测高分子加权蛋白。这是硝酸纤维素和PVDF膜之间的差异。
图片提供:
1。”Western blot transfer” By Bensaccount at English Wikipedia(CC由3.0)通过下议院维基梅迪亚
参考:
1。Luque-Garcia, Jose L., Ge Zhou, Tung-Tien Sun, and Thomas A. Neubert. “Use of Nitrocellulose Membranes for Protein Characterization by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry.” Analytical chemistry. U.S. National Library of Medicine, 15 July 2006. Web. 28 Mar. 2017
2.“分子标记和植物生物技术。”Google书籍。N.P.,N.D。网络。2017年3月28日
3. 2013年4月8日的Monday Tweet。“膜上的魔术:明智地选择最佳蛋白质印迹结果。”生物组合。N.P.,2013年4月8日。2017年3月28日
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