Erratum: Detergent Lysis of Tissue Culture Cells for Immunoprecipitation
Cold Spring Harb Protoc 2017; doi: 10.1101/pdb.prot098558
When the above protocol was published, multiple reagents in the Nonidet P-40 Lysis Buffer and RIPA Lysis Buffer recipes and/or their volumes and concentrations were incorrect. The publisher apologizes for any confusion related to these errors, which appear to have been inadvertently introduced when the protocol was formatted for publication. The PDF version of the protocol and the HTML versions of the Nonidet P-40 Lysis Buffer (doi: 10.1101/pdb.rec101410) and RIPA Lysis Buffer (10.1101/pdb.rec101428) recipes have been replaced with the corrected recipes shown below. The originally published versions of the recipes are also included.
CORRECTED RECIPES
Nonidet P-40 Lysis Buffer
| Reagent | Final concentration |
|---|---|
| NaCl | 150 mm |
| Nonidet P-40 | 1% |
| Tris (pH 8.0) | 50 mm |
| Prepared Nonidet P-40 lysis buffer should be aliquoted and stored at −20°C. Add protease and/or phosphatase inhibitors to a thawed aliquot before immediate use. Discard and do not freeze again. | |
RIPA Lysis Buffer
| Reagent | Final concentration |
|---|---|
| NaCl | 150 mm |
| Nonidet P-40 | 1% |
| DOC | 0.5% |
| SDS | 0.1% |
| Tris (pH 7.4) | 50 mm |
| Prepared RIPA buffer should be aliquoted and stored at −20°C. Add protease and/or phosphatase inhibitors to a thawed aliquot before immediate use. Discard and do not freeze again. One milliliter of buffer is sufficient to lyse approximately 5 million cells. | |
RECIPES AS ORIGINALLY PUBLISHED
Nonidet P-40 Lysis Buffer
| Reagent | Volume per 10 ml of solution (v/v) | Final concentration |
|---|---|---|
| NaCl (150 mm) | 0.66 ml | 10 mm |
| Nonidet P-40 (1%) | 0.1 ml | 1% |
| Tris (50 mm, pH 7.4) | 2 ml | 10 mm |
| MgCl2 (10 mm) | 3 ml | 3 mm |
| ddH2O | Add to 10 ml | |
| Prepared Nonidet P-40 lysis buffer should be aliquoted and stored at −20°C. Add protease and/or phosphatase inhibitors to a thawed aliquot before immediate use. Discard and do not freeze again. | ||
RIPA Lysis Buffer
| Reagent | Volume per 10 ml of solution (v/v) | Final concentration |
|---|---|---|
| NaCl (1 M) | 1.5 ml | 150 mm |
| Nonidet P-40 (1%) | 0.1 ml | 1% |
| Sodium deoxycholate (DOC) (0.5%) | 0.05 ml | 0.5% |
| SDS (0.1%) | 0.01 ml | 0.1% |
| Tris (50 mm, pH 7.4) | 5 ml | 25 mm |
| ddH2O | Add to 10 ml | |
| Prepared RIPA buffer should be aliquoted and stored at −20°C. Add protease and/or phosphatase inhibitors to a thawed aliquot before immediate use. Discard and do not freeze again. This amount is sufficient for approximately 5 million cells per 1 ml of reagent. | ||
