蛋白质相关实验技术操作全攻略

1、Western blotting:

Western blotting using ECL reagent
used by：Laboratory of P.J. Hansen,Dept. of Animal Sciences, University of Florida
Kamps's Western Blotting Protocol
used by：Sefton Lab, Salk Institute
Dry Transfer 干法蛋白转膜
used by：The Preuss Lab,The Division of Biological Sciences,The University of Chicago
Western Blot of TBP from TBP-GST bacteria
used by：Steven Finkbeiner, Departments of Neurology and Physiology, UCSF
Western Blot/Anti-P-CREB
used by：Steven Finkbeiner, Departments of Neurology and Physiology, UCSF
Western blotting protocol for 1C2, 3B5H10, and 4C8 Antibodies
used by：Steven Finkbeiner, Departments of Neurology and Physiology, UCSF
Protocol for anti-HA antibody Western Blotting
used by：Steven Finkbeiner, Departments of Neurology and Physiology, UCSF
Probing and Stripping of Western Blot
used by：Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute
Western Blotting Protocol
used by：Mirmira Laboratory at the University of Virginia
Western Blotting and Immunostaining
used by：Chazin Lab, Center for Structural Biology, Vanderbilt University
Western Blots
used by：Lab of Dr. Mark Barton Frank, Oklahoma Medical Research Foundation
Western Blotting - Antibodies
used by：The Minion Lab, College of Veterinary Medicine at Iowa State University
Western Blotting
used by：Cepko/Tabin Lab, Harvard University
Carbonate Solution For Western Blot
used by：Dr. DE Koshland, Carnegie Institution of Washington
Western Blot with BSA
used by：Dr. DE Koshland, Carnegie Institution of Washington
METHOD for Western Blots
used by：Dr. DE Koshland, Carnegie Institution of Washington
Western Blots
used by：Michael Koelle's Lab,Department of Molecular Biophysics and Biochemistry
Yale University

2、蛋白分离/提取/浓缩

Lysis Buffers
used by：Steven Finkbeiner, Departments of Neurology and Physiology, UCSF
Yeast Protein Isolation "Yaffe-Schatz"
used by：Amberg Lab ,Upstate Medical University
Guanidine Hydrochloride Purification of Proteins From SDS PAGE
used by：Gottschling Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute
Yeast protein prep for SDS PAGE and western (rapid)
used by：Gottschling Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute
Yeast protein prep for SDS PAGE and western (glass bead)
used by：Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute
Purification of TFIIIC from yeast
used by：Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute
Purification of TFIIA from E. coli
used by：Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute
TCA protein precipitation
used by：Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute
Purification of MLCK ( Myosin Light Chain Kinase )from Rabbit Skeletal Muscle肌球蛋白轻链激酶
used by：Stull Lab,University of Texas, Southwestern Medical Center
Purification of Calmodulin from Bovine Brain
used by：Stull Lab,University of Texas, Southwestern Medical Center
Myosin Light Chain Preparation from Skeletal and Cardiac Muscle
used by：Stull Lab,University of Texas, Southwestern Medical Center
Purifying Protein from Inclusion Bodies
used by：Chazin Lab, Center for Structural Biology, Vanderbilt University
Protocol for Protein Extraction from Plant
used by：The University of Nebraska-Lincoln Protein Core Facility (PCF)
Taq Polymerase Purification
used by：Cepko/Tabin Lab, Harvard University
Yeast Protein Prep (No Boil)
used by：Dr. DE Koshland, Carnegie Institution of Washington
Yeast Protein Prep (Boil)
used by：Dr. DE Koshland, Carnegie Institution of Washington
GST Fusion Protein Prep
used by：Vesicle Trafficking, Stanford University

3、蛋白质检测分析：

AMIDO BLACK STAIN
used by：Hancock Laboratory Methods,Department of Microbiology and Immunology,
University of British Columbia, British Columbia, Canada
James Hardwick's angiotensin assay protocol
used by：Sefton Lab,Salk Institute
Phosphoamino acid analysis:Mark Kamps's method
used by：Sefton Lab,Salk Institute
ULTIMATE HIS-UB ASSAY FOR
used by：the Tansey Lab at Cold Spring Harbor Laboratory
ULTIMATE HIS-UB ASSAY FOR YEAST
used by：the Tansey Lab at Cold Spring Harbor Laboratory
Protocol for Beta-Hexosaminidase RBL-2H3 Secretion Assay
used by：meyer lab,Stanford University
Measurement of Myosin Regulatory Light Chain (RLC) Phosphorylation
used by：Stull Lab,University of Texas, Southwestern Medical Center
Bicinchoninic Acid (BCA) Protein Assay
used by：David R. Caprette, Rice University
Biuret Protein Assay
used by：David R. Caprette, Rice University
Bradford protein assay
used by：David R. Caprette, Rice University
Hartree-Lowry and Modified Lowry Protein Assays
used by：David R. Caprette, Rice University
Colorimetric Assays
used by：David R. Caprette, Rice University
Kd measurement, fitting, calculation, and simulation
used by：Chazin Lab, Center for Structural Biology, Vanderbilt University

4、蛋白标记修饰

DSP Crosslinking
used by：Peter Novick Lab, Department of Cell Biology Yale University School of Medicine
Labeling Gc-Globulin with Carboxyfluorescein Succinimidyl Ester
used by：Yu-Li Wang Lab,University of massachusetts Medical School(UMASS)
Labeling Tubulin with Tetramethylrhodamine Succinimidyl Ester
used by：Yu-Li Wang Lab,University of massachusetts Medical School(UMASS)
Labeling Gizzard Vinculin with Tetramethylrhodamine Iodoacetamide
used by：Yu-Li Wang Lab,University of massachusetts Medical School(UMASS)
Labeling Gizzard Alpha-actinin with Tetramethylrhodamine Iodoacetamide
used by：Yu-Li Wang Lab,University of massachusetts Medical School(UMASS)
Labeling Gizzard Myosin with Tetramethylrhodamine Iodoacetamide (primarily 17kD MLC)
used by：Yu-Li Wang Lab,University of massachusetts Medical School(UMASS)
Labeling Gizzard Myosin with Tetramethyorhodamine Iodoacetamide (MHC and 17kD MLC)
used by：Yu-Li Wang Lab,University of massachusetts Medical School(UMASS)
Labeling Muscle Actin with N-(1-pyrene) Iodoacetamide
used by：Yu-Li Wang Lab,University of massachusetts Medical School(UMASS)
Labeling Muscle Actin with Tetramethylrhodamine Iodoacetamide
used by：Yu-Li Wang Lab,University of massachusetts Medical School(UMASS)
Labeling Muscle Actin with Carboxyfluorescein Succinimidyle Ester
used by：Yu-Li Wang Lab,University of massachusetts Medical School(UMASS)
Labeling Muscle Actin with 5-Iodoacetamidofluorescein
used by：Yu-Li Wang Lab,University of massachusetts Medical School(UMASS)
ultimate mammalian cell pulse-chase
used by：the Tansey Lab at Cold Spring Harbor Laboratory
ultimate yeast pulse-chase
used by：the Tansey Lab at Cold Spring Harbor Laboratory
Gluteraldehyde Conjugation of Oligopeptides to Proteins
used by：Lab of Dr. Mark Barton Frank, Oklahoma Medical Research Foundation
Fluorescein labeling of proteins
used by：David Chambers,Salk institute
Biosynthetic labeling
used by：Sefton Lab, Salk Institute , San Diego, California

5、免疫沉淀(IP/CoIP/ChIP)

Immunoprecipitation buffers and protocols
used by：Steven Finkbeiner, Departments of Neurology and Physiology, UCSF
Cell Lysis/Western/IP
used by：Steven Finkbeiner, Departments of Neurology and Physiology, UCSF
Amplification and Labelling with Cy dyes
used by：Yale Genome Analysis Center, Yale University
Chromatin Immunoprecipitation from Mammalian Cell Extracts
used by：Yale Genome Analysis Center, Yale University
Chromatin Immunoprecipitation from Yeast Whole Cell Extracts
used by：Yale Genome Analysis Center, Yale University
Immunoprecipitation
used by：Hancock Laboratory Methods. Department of Microbiology and Immunology,
University of British Columbia, British Columbia, Canada
Ultimate yeast denaturing IP
used by：the Tansey Lab at Cold Spring Harbor Laboratory
ULTIMATE FREEZE-THAW LYSIS FOR
used by：the Tansey Lab at Cold Spring Harbor Laboratory
Ultimate mammalian nondenaturing IP
used by：the Tansey Lab at Cold Spring Harbor Laboratory
Ultimate mammalian denaturing IP
used by：the Tansey Lab at Cold Spring Harbor Laboratory
Ultimate yeast ChIP assay
used by：the Tansey Lab at Cold Spring Harbor Laborat ory
Chromatin Immunoprecipitation Assay and PCR
used by：Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute
Yeast Chromatin Immunoprecipitation (ChIP)
used by：Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute
Mirmira Lab ChIP Protocol
used by：Mirmira Laboratory at the University of Virginia
ChIP Assay Protocol
used by：Mirmira Laboratory at the University of Virginia
Immunoprecipitation and Immune Complex kinase assay
used by：Sefton Lab, Salk Institute
General Principles of Immunoprecipitation
used by：Sefton Lab, Salk Institute

6、融合蛋白表达及纯化

Use of B-PER lysis reagent to purify recombinant proteins tagged with His6 or maltose binding protein
used by：Smith Lab,Division of Biological Sciences,University of Missouri
Bacterial Expression of Kinesin Motors
used by：Liz Greene and Steve Henikoff
Purification of recombinant sBRF M166L
used by：Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute
Protein Expression and Purification of RPA70-AB
used by：Chazin Lab, Center for Structural Biology, Vanderbilt University
Co-Expression and purification of human DNA primase p49-p58 subunits引发酶p49-p58亚基共表达纯化方法
used by：Chazin Lab, Center for Structural Biology, Vanderbilt University
Protein Expression and Purification of MBP-T antigen MBP-T抗原表达纯化
used by：Chazin Lab, Center for Structural Biology, Vanderbilt University
Perspectives on Baculovirus Expression Systems表达
used by：Lab of Dr. Mark Barton Frank, Oklahoma Medical Research Foundation
Expression & Purification of His Tagged Proteins in E. coli
used by：Cepko/Tabin Lab, Harvard University

7、相互作用分析

Amplifying a large phage-display library without losing diversity
used by：Smith Lab,Division of Biological Sciences,University of Missouri
Protein Interaction Analysis Using an IASYS Biosensor
used by：Kitto Lab, The University of Texas at Austin
Yeast Two-Hybrid Screen with Library and Bait
used by：Mirmira Laboratory at the University of Virginia

8、蛋白电泳/双向电泳

SDS-PAGE
used by：Steven Finkbeiner, Departments of Neurology and Physiology, UCSF
Gel Drying干胶
used by：Steven Finkbeiner, Departments of Neurology and Physiology, UCSF
NuPAGE Gel Electrophoresis 蛋白电泳
used by：Kitto Lab, The University of Texas at Austin
Isoelectric Focussing of Membrane Proteins by Slab Gel Method
used by：Hancock Laboratory Methods,Department of Microbiology and Immunology,
University of British Columbia, British Columbia, Canada
Silver Staining SDS PAGE Gels
used by：Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute
E. coli total protein for SDS PAGE
used by：Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute
Comassie Blue Protein gel stain
used by：Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute
Coomassie & Silver Staining of Polyacrylamide Gels
used by：Lab of Dr. Mark Barton Frank, Oklahoma Medical Research Foundation
2-D Polyacrylamide Gel Electrophoresis
used by：Molecular Profiling Initiative, National Cancer Institute(NCI)
Silver Staining of SDS-PAGE Gels
used by：The Minion Lab, College of Veterinary Medicine at Iowa State University
SDS-PAGE Gels
used by：The Minion Lab, College of Veterinary Medicine at Iowa State University
CuCl2 Staining of SDS-PAGE Gels
used by：Cepko/Tabin Lab, Harvard University
Electroelution of Proteins From SDS-PAGE Gels
used by：Drs. C Cepko and C Tabin, Harvard University
Silver Stain of Protein Gels
used by：Dr. DE Koshland, Carnegie Institution of Washington
SDS-PAGE （BIO-RAD
used by：Dr. DE Koshland, Carnegie Institution of Washington
SDS-PAGE
used by：Michael Koelle's Lab,Department of Molecular Biophysics and Biochemistry
Yale University

9、质谱分析

Polycrystalline thin films质谱样品准备
used by：Prowl Lab,Rockefeller University
Seeded films (Vorm-Roepstorff)
used by：Prowl Lab,Rockefeller University
Dried droplet
used by：Prowl Lab,Rockefeller University
In-gel Tryptic Digest for Protein ID by Mass Spectrometry
used by：Mitshison Lab, Department of Systems Biology, Harvard Medical School

10、crystallization

Polycrystalline thin films
used by：PROWL, Rockefeller University
Slow crystallization
used by：PROWL, Rockefeller University
Crystallization of Kinesin Family Motor Proteins驱动蛋白家族的结晶条件和晶体参数
used by：Liz Greene and Steve Henikoff
Crystallization Trials
used by：Liz Greene and Steve Henikoff

11、in vitro protein synthesis

Protein Syntheses in Cell Free Systems
used by：The Laboratory of William Heidcamp at Gustavus Adolphus University

12、蛋白纯化/层析

6xHis-tagged protein purification using Qiagen Ni-NTA Column under Native Conditions
used by：Sugden lab,McArdle Laboratory for Cancer Research, University of Wisconsin-Madison Medical School
Purification of dnEBNA-1/Soft from E. coli BL21 LysS
used by：Sugden lab,McArdle Laboratory for Cancer Research, University of Wisconsin-Madison Medical School
Purification of GST fusion proteins in E.coli GST融合蛋白纯化（方法四）筛选表达株
used by：Sugden lab,McArdle Laboratory for Cancer Research, University of Wisconsin-Madison Medical School
Purification of GST fusion proteins in E.coli GST融合蛋白纯化，方法三，纯化小量蛋白
used by：Sugden lab,McArdle Laboratory for Cancer Research, University of Wisconsin-Madison Medical School
Purification of GST fusion proteins in E.coli GST融合蛋白纯化，方法二
used by：Sugden lab,McArdle Laboratory for Cancer Research, University of Wisconsin-Madison Medical School
Purification of GST fusion proteins in E.coli GST融合蛋白纯化，方法一
used by：Sugden lab,McArdle Laboratory for Cancer Research, University of Wisconsin-Madison Medical School
Purification of 6xHis epitope tagged proteins by Ni-NTA-Agarose His标签蛋白纯化
used by：Sugden lab,McArdle Laboratory for Cancer Research, University of Wisconsin-Madison Medical School
Fusion Protein Isolation 融合蛋白分离纯化
used by：Peter Novick Lab, Department of Cell Biology Yale University School of Medicine
Affinity Column Preparation 亲和层析柱制备
used by：Peter Novick Lab, Department of Cell Biology Yale University School of Medicine
Preparation of Affinity Column制备亲和层析柱
used by：Steven Finkbeiner, Departments of Neurology and Physiology, UCSF
Column Buffers
used by：Steven Finkbeiner, Departments of Neurology and Physiology, UCSF
13、蛋白定量

Glutathione - a microassay for determining glutathione content in cells 检测细胞谷胱苷肽含量
used by：Laboratory of P.J. Hansen,Dept. of Animal Sciences, University of Florida
Bradford Protein Assay - a microassay for determining protein content in a 96-well microtiter plate format
used by：Laboratory of P.J. Hansen,Dept. of Animal Sciences, University of Florida
Lowry Protein Assay
used by：Laboratory of P.J. Hansen,Dept. of Animal Sciences, University of Florida
Bradford Assay
used by：Kitto Lab, The University of Texas at Austin

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