• 我要登录|
  • 免费注册
    |
  • 我的丁香通
    • 企业机构:
    • 成为企业机构
    • 个人用户:
    • 个人中心
  • 移动端
    移动端
丁香通 logo丁香实验_LOGO
搜实验

    大家都在搜

      大家都在搜

        0 人通过求购买到了急需的产品
        免费发布求购
        发布求购
        点赞
        收藏
        wx-share
        分享

        【资源】Electrophoresis 知识

        丁香园论坛

        935
        Electrophoresis其实并不简单
        我们要学的还很多
        Chemical Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 334
        What Is the Safest Approach to Working with
        Acrylamide? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 334
        What Are the Symptoms of Acrylamide Poisoning? . . . . . . 335
        What Is the Medical Response to Accidental Acrylamide
        Exposure? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 335
        How Can You Dispose of Excess, Unusable Acrylamide? . . 335
        What Is the Shelf Life of Acrylamide and Acrylamide
        Solutions? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 336
        Electrical Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 336
        What Are the Requirements for a Safe Work Area? . . . . . . 336
        What Are the Requirements for Safe Equipment in Good
        Working Order? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 337
        Polyacrylamide (PAGE) Gels—Before Selecting a Gel:
        Getting the Best Results for Your Purpose . . . . . . . . . . . . . . . . 337
        What Is the Mechanism of Acrylamide Polymerization? . . . 338
        What Other Crosslinkers Are Available, and When
        Should They Be Used? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 338
        How Do You Control Pore Size? . . . . . . . . . . . . . . . . . . . . . . 339
        How Do You Calculate %T and %C? . . . . . . . . . . . . . . . . . . . 341
        Why Should You Overlay the Gel? What Should You Use
        for an Overlay? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 341
        Regarding Reproducible Polymerization, What Practices
        Will Ensure That Your Bands Run the Same Way Every
        Time? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 341
        What Catalyst Concentration Should You Use? . . . . . . . . . . 343
        What Is the Importance of Reagent Purity on Protein
        Electrophoresis and Staining? . . . . . . . . . . . . . . . . . . . . . . . 343
        Which Gel Should You Use? SDS-PAGE, Native PAGE or
        Isoelectric Focusing? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 345
        Will Your SDS Gel Accurately Indicate the Molecular
        Weight of Your Proteins? . . . . . . . . . . . . . . . . . . . . . . . . . . . 345
        Should You Use a Straight % Gel or a Gradient Gel? . . . . . 345
        What Issues Are Relevant for Isoelectric Focusing? . . . . . . 346
        How Can You Resolve Proteins between Approximately
        300 and 1000kDa? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 347
        What Issues Are Critical for Successful Native PAGE? . . . . . . 348
        Sample Solubility . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 348
        Location of Band of Interest . . . . . . . . . . . . . . . . . . . . . . . . . 348
        How Can You Be Sure That Your Proteins Have Sufficient
        Negative Charge to Migrate Well into a Native PAGE
        Gel? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 348
        Buffer Systems for Native PAGE . . . . . . . . . . . . . . . . . . . . . . . 349
        What Can Go Wrong with the Performance of a
        Discontinuous Buffer System? . . . . . . . . . . . . . . . . . . . . . . . . . . 349
        What Buffer System Should You Use for Peptide
        Electrophoresis? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 350
        Power Issues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 350
        Constant Current or Constant Voltage—When and
        Why? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 351
        Why Are Nucleic Acids Almost Always Separated via
        Constant Voltage? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 352
        Why Are Sequencing Gels Electrophoresed under
        Constant Power? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 352
        Should You Run Two Sequencing Cells off the Same Power
        Supply under Constant Power? . . . . . . . . . . . . . . . . . . . . . 352
        Improving Resolution and Clarity of Protein Gels . . . . . . . . . 353
        How Can You Generate Reproducible Gels with Perfect
        Bands Every Time? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 353
        Sample Preparation—Problems with Protein Samples . . . . . . 353
        What Procedures and Strategies Should Be Used to
        Optimize Protein Sample Preparation? . . . . . . . . . . . . . . . 353
        Is the Problem Caused by Sample Preparation or by the
        Electrophoresis? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 354
        Is the Problem Caused by the Sample or the Sample
        Buffer? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 354
        How Do You Choose a Detergent for IEF or Native
        PAGE? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 354
        What Other Additives Can Be Used to Enhance Protein
        Solubility? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 355
        Agarose Electrophoresis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 355
        What Is Agarose? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 355
        What Is Electroendosmosis (-Mr or EEO)? . . . . . . . . . . . . . . 355
        Are Double-Stranded Markers Appropriate for Sizing
        Large Single-Stranded (Not Oligonucleotide)
        DNA? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 356
        What Causes Nucleic Acids to Migrate at Unexpected
        Migration Rates? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 356
        What Causes Commercial Preparations of Nucleic Acid
        Markers to Smear? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 356
        What Causes Fuzzy Bands? . . . . . . . . . . . . . . . . . . . . . . . . . . . 357
        Elution of Nucleic Acids and Proteins from Gels . . . . . . . . . . . 357
        Detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 357
        What Should You Consider before Selecting a Stain? . . . . . . 357
        Will the Choice of Stain Affect a Downstream
        Application? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359
        Is Special Equipment Needed to View the Stain? . . . . . . . . . 361
        How Much Time Is Required for the Various
        Stains? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 361
        What If You Need to Quantify Your Stained Protein? . . . . . 361
        What Causes High Background Staining? . . . . . . . . . . . . . . . 362
        Will the Presence of Stain on Western-Blotted Proteins
        Interfere with Subsequent Hybridization or Antibody
        Detection Reactions? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 363
        Does Ethidium Bromide Interfere with the Common
        Enzymatic Manipulation of Nucleic Acids? . . . . . . . . . . . . . 363
        Standardizing Your Gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 363
        What Factors Should Be Considered before Selecting a
        Molecular Weight Marker? . . . . . . . . . . . . . . . . . . . . . . . . . . 363
        Are Double-Stranded Markers Appropriate for Sizing
        Large (Not Oligonucleotide) Single-Stranded DNA? If
        Not, Which Markers Are Recommended? . . . . . . . . . . . . 364
        Can a Pre-stained Standard Be Applied to Determine the
        Molecular Weight of an Unknown Protein? . . . . . . . . . . . 364
        How Do You Determine Molecular Weight on a
        Western Blot? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 365
        What Are the Options for Determining pI and Molecular
        Weight on a 2-D Gel? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 365
        How Do You Measure the pH Gradient of a Tube IEF Gel
        or an IPG Gel? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 366
        Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 368
        What Is This Band Going All the Way across a Silver-
        Stained Gel, between Approximately 55 and
        65kDa? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 368
        How Can You Stop the Buffer Leaking from the Upper
        Chamber of a Vertical Slab Cell? . . . . . . . . . . . . . . . . . . . . . 368
        Bibliography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 368
        Appendix A: Procedure for Degassing Acrylamide Gel
        Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 371
        ad image
        提问
        扫一扫
        丁香实验小程序二维码
        实验小助手
        丁香实验公众号二维码
        扫码领资料
        反馈
        TOP
        打开小程序