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Antibody Explorer
Nondenaturing Protein Immunoprecipitation
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Nondenaturing Protein Immunoprecipitation from Mammalian Cells
William P. Tansey
ABSTRACT
This protocol describes a nondenaturing immunoprecipitation (IP) for mammalian cells. Where possible, we prefer to use denaturing IPs to recover labeled proteins from pulse-chase experiments. However, the nondenaturing protocol is useful when one wishes to separate soluble from insoluble proteins, or when the antibody being used recognizes a native epitope.
Products Available for this Protocol
| Protocol Material |
Product No. |
Description |
Add to Cart |
| Buffers, Solutions, and Reagents |
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| Laemmli (SDS-PAGE) sample buffer (2X) |
S3401 |
Sample Buffer, Laemmli 2× Concentrate |
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| Protein A Sepharose (50%, pre-swollen) |
P3391 |
Protein A-Sepharose® from Staphylococcus aureus |
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| Protein G Sepharose (50%, pre-swollen) |
P3296 |
Protein G Sepharose®, Fast Flow, recombinant, expressed in Escherichia coli, aqueous ethanol suspension |
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| NaCl |
S3014 |
Sodium chloride, for molecular biology |
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| Pefabloc |
76307 |
Pefabloc® SC |
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| NP-40 |
NP40S |
Tergitol® solution, Type NP-40, 70% in H2O |
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| Deoxycholate (DOC) |
D6875 |
21-Hydroxyprogesterone |
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| SDS (10%) |
L4390 |
Sodium dodecyl sulfate, for molecular biology |
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| Tris-Cl |
T5941 |
Trizma® hydrochloride, for molecular biology, Biotechnology Performance Certified |
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| Dithiothreitol (DTT) |
D9779 |
DL-Dithiothreitol, for molecular biology |
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| Bromophenol blue |
B0126 |
Bromophenol Blue |
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| Aprotinin |
A6103 |
Aprotinin from bovine lung, recombinant, expressed in Nicotiana |
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| Pepstatin |
P5318 |
Pepstatin A |
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| Leupeptin |
L9783 |
Leupeptin hydrochloride |
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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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