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Protein Biology Learning Center

Learn about protein analysis methods and technologies

从这里开始学习或进一步提升您的蛋白质生物学知识

蛋白质生物学不仅包含研究内容,也包含了研究所需的工具。即基于蛋白质结构及功能的研究,和利用抗体、蛋白及多肽等作为纯化、检测和分析生物系统的工具。有些蛋白分析方法(如western blotting和ELISA等)已作为实验室的常规实验方法使用了多年。其它方法如定量蛋白质谱的实验方法则是相对较新的实验技术,并且正在快速发展中。此外,全新的工具和产品也在持续不断地被研发出来,它们共同推进了所有蛋白质生物学研究领域的发展.

本学习中心通过提供蛋白质研究的一些关键应用,使科研工作者能够轻松的访海量蛋白质研究资源,以便学习和了解蛋白质分析方法及产品.
Protein biology guided learning

视频演示了western blot流程中的每一个步骤,包括SDS-PAGE、转膜、封闭、使用抗体进行探针检测、化学发光检测及印迹剥离等步骤.

回顾质谱的发展历史、应用领域及方法论等,包括离子化(ESA,MALDI)、质量分析仪(TOF,四级杆,离子阱),离子检测器、MS/MS、LC-MS/MS及样品制备等内容.

该指南提供了如何选择适合底物,并根据底物选择起始的一抗和二抗稀释浓度所需的相关信息.

副主题

Protein Isolation and Purification

Learn about methods and technologies for efficient cell lysis, protein extraction and fractionation, targeted inhibition of unwanted protease and phosphatase activity, and convenient devices and high performance resins for the purification and clean-up of proteins and antibodies for downstream applications.

  • Cell lysis
  • Protein extraction
  • Protease & phosphatase inhibitors
  • Dialysis
  • Desalting
  • Sample clean-up
  • Affinity purification
  • Resins, beads, supports
  • Ligand immobilization
  • Immunoprecipitation (IP) 
  • Pull-down
  • Technical handbooks
  • Protein methods articles
  • Product application notes
  • Support center resources
  • Protocols
  • Product selection guides

Protein Gel Electrophoresis

Learn about methods and technologies for separation and analysis of proteins by polyacrylamide gel electrophoresis (PAGE). Understand gel types (pouring, precast, gradient, 1D, 2D, etc.), gel and running buffer systems, sample preparation and loading, molecular weight markers (protein ladders), and gel stains.

  • Precast gels
  • Pouring gels
  • Gradient gels
  • Buffer systems
  • Gel tanks
  • Sample loading
  • MW markers & ladders
  • Gel stains
  • Technical handbooks
  • Protein methods articles 
  • Product application notes
  • Support center resources
  • Protocols
  • Product selection guides

Western Blotting

Learn about methods and technologies for western blot detection and imaging. Understand the reagents and instruments used for the efficient membrane transfer and detection of proteins, including wash and blocking buffers, secondary antibody conjugates, and chemiluminescent, colorimetric or fluorescent detection substrates.

  • Tank vs. semi-dry transfer 
  • Blotting membranes
  • Nitrocellulose vs. PVDF
  • Blocking buffers
  • Washing buffers 
  • Detection probes
  • Antibodies
  • Enzymes & substrates
  • Chemiluminescent vs. fluorescent detection
  • Imaging
  • Technical handbooks
  • Protein methods articles
  • Product application notes
  • Support center resources
  • Protocols 
  • Product selection guides

Protein Assays and Analysis

Learn about methods and technologies for identifying and measuring proteins. Understand protein quantitation reagents, ELISA and other immunoassays, protein interaction and activity assays, enzyme reporter assays, protein microarrays, and immunoprecipitation and pull-down tools.

  • BCA, Coomassie dye, & fluorescent protein assays
  • BSA standards
  • A280
  • Immunoprecipitation (IP)
  • Pull-down
  • ELISA 
  • Immunoassays
  • Blocking buffers
  • Multiplex
  • Microarrays
  • Enzyme reporter assays
  • Activity assays 
  • Technical handbooks
  • Protein methods articles
  • Product application notes
  • Support center resources
  • Protocols
  • Product selection guides

Protein Mass Spectrometry

Learn about methods and technologies for peptide and protein analysis by high performance liquid chromatography and mass spectrometry (LC/MS). Understand sample prep tools, mass and isotope labeling strategies for quantitative proteomics, and the basics of MS data and analysis.

  • Proteomics
  • Cell lysis
  • Protein enrichment
  • Trypsin digestion
  • Peptide clean-up
  • C18
  • Sample prep 
  • Isobaric mass tagging
  • Stable isotope labeling
  • Metabolic labeling
  • Tandem mass spec
  • MS/MS
  • LC/MS
  • MALDI
  • Orbitrap
  • Technical handbooks
  • Protein methods articles
  • Product application notes
  • Support center resources
  • Protocols
  • Product selection guides

Protein Modification and Crosslinking

Learn about methods and technologies for protein labeling, modification, and crosslinking. Understand the chemical functional groups and reaction chemistries required to crosslink and conjugate proteins to one another; to label proteins with biotin, PEG, or fluorescent dyes; or to reduce, cleave, and fragment proteins for use in assays.

  • Crosslinking
  • Crosslinker chemistry
  • Conjugation reagents 
  • Bioconjugates
  • Covalent attachment
  • Functional groups
  • Reactive moieties
  • Amino acids 
  • Amines
  • Sulfhydryls
  • Carboxyls
  • NHS esters
  • Maleimides
  • EDC
  • EDAC
  • Molecular probes 
  • Fluorescent dyes
  • Biotinylation
  • PEGylation
  • Technical handbooks
  • Protein methods articles
  • Product application notes
  • Support center resources
  • Protocols
  • Product selection guides

Antibodies and Proteins

Learn about methods and technologies for preparing and using antibodies and recombinant proteins in protein analysis techniques. Understand how specific primary and secondary antibodies are produced, screened, purified, and modified to enable their use in a myriad of protein detection (identification and measurement) applications.

  • Primary antibodies
  • Secondary antibodies
  • Antibody production
  • Immunogen preparation
  • Antibody purification
  • Antibody labeling 
  • Western blotting
  • ELISA
  • Multiplex immunoassays
  • Microarrays
  • Immunoprecipitation
  • Technical handbooks 
  • Protein methods articles
  • Product application notes
  • Support center resources
  • Protocols
  • Product selection guides

Resource library features

Methods article (videos)

Videos that demonstrate each step in the western blot workflow, including SDS-PAGE, transfer, blocking, probing with antibodies, chemiluminescent detection, and stripping the blot.

Methods article

A review of the enzyme-linked immunsorbant assay (ELISA), including formats (sandwich), detection methods (direct, indirect), plate coating techniques, antibodies and other probes, buffers, and kits.

Methods article

Strategies for proteomic analyses, including those for relative quantitation (SILAC, ICAT, ICPL and isobaric tags) and for absolute quantitation (via SRM), and both labeled and label-free techniques.