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The following Cold Spring Harbor Laboratory Press protocols are brought to you by BioSupplyNet in a partnership with Sigma-Aldrich. A list of products is available to use in each protocol.
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Selected Protocol from "Proteins and Proteomics: A Laboratory Manual," Richard J. Simpson.
View the protocol
"Analysis of the Topology of Protein Complexes Using Cross-Linking and Mass Spectrometry" at BioSupplynet.com
Juri Rappsilber and Matthias Mann
ABSTRACT
This protocol is designed to allow identification of spatial relationships between proteins within a wide variety of multiprotein complexes. Details for optimizing cross-linking of proteins within multiprotein complexes are provided. Following
the cross-linking reaction, the proteins are separated by one-dimensional
PAGE, the cross-linked proteins are isolated from the gel, and the individual members of the cross-linked complexes are identified by mass spectrometry. The protocol can be used to analyze many protein complexes isolated by any purification technique, provided the protein complexes remain in their native configuration.
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Products Available for this Protocol
| Protocol Material Description |
Product # |
Product Name |
Add to Cart |
| 20 mM PIPES (pH 7.0) |
P1851 |
PIPES, Biotechnology Performance Certified, cell culture tested, ≥99% (titration) |
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| BS³ (bis[sulfosuccinimidyl]suberate) |
S5799 |
Suberic acid bis(3-sulfo-N-hydroxysuccinimide ester) sodium salt |
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| Dithiothreitol (DTT) |
D9779 |
DL-Dithiothreitol, for molecular biology, ≥98% (TLC), ≥99% (titration)
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| Tris-Cl (pH 7.8) |
93313 |
Trizma® hydrochloride buffer solution, BioChemika Ultra, for molecular biology, pH 7.4 |
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| Dimethylsulfoxide (DMSO) (HPLC grade, if possible) |
34869 |
Dimethyl sulfoxide, CHROMASOLV® Plus, for HPLC, ≥99.7% |
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| EDC (1-ethyl-3-[3-dimethylaminopropyl]carbodiimide) |
165344 |
1-[3-(Dimethylamino)propyl]-3-ethylcarbodiimide methiodide |
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| SMCC (succinimidyl-4-[N-maleimidomethyl]cyclohexane-1-carboxylate) |
M6035 |
4-(N-Maleimidomethyl)cyclohexane-1-carboxylic acid 3-sulfo-N-hydroxysuccinimide ester sodium salt |
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| SDS |
L4390 |
Sodium dodecyl sulfate, for molecular biology, ≥98.5% (GC) |
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| Bromophenol blue |
B5525 |
Bromophenol Blue sodium salt for electrophoresis |
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| Glycerol |
G5516 |
Glycerol, for molecular biology, ≥99% |
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| Acrylamide solution |
A4058 |
Acrylamide solution, 40%, for electrophoresis, sterile-filtered |
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| Protein molecular weight marker, prestained |
View the entire line of protein molecular weight markers |
| Reagents for protein staining |
View the entire line staining reagents for proteins |
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Product Association Disclaimer: The Sigma-Aldrich products listed for this specific protocol were selected either to match or to supplement the products listed within the actual protocol. The products/reagents from Sigma-Aldrich have been qualified for usage, but may not have been validated for this specific application. Please refer to the detailed product description on the usage of specific products of interest.
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