Hi all,
I am looking into purchasing the Thermo Scientific Pierce Modified Lowry Protein Assay Kit (Catalog number: 23240) to quantify total protein amount in mammalian cell lysates. I lyse my cells in RIPA buffer (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, 0.1% SDS) and am wondering if this buffer is compatible with the kit.
I appreciate any advice you can offer! Thank you!
Yes, RIPA buffer is compatible with the Thermo Scientific Pierce Modified Lowry protein quantification assay.
Hello Melissa Rogers
I agree with İsmail Emir Akyildiz . BCA protein assay is the right choice. You may go through Table 1 of the attached link provided below that summarizes compatible substances for several popular protein assays. The table serves as a guide for assessing which protein assay might be most effective with a given sample type. From the table it is quite well understood that with RIPA buffer you should go with BCA protein assay.
https://assets.fishersci.com/TFS-Assets/LSG/Application-Notes/TR0068-Protein-assay-compatibility.pdf
I would suggest that instead of purchasing the Thermo Scientific Pierce Modified Lowry Protein Assay Kit (Catalog number: 23240), you should go for the Thermo Scientific Pierce™ BCA Protein Assay Kit (Cat. No. 23225) or (Cat. No. 23227). RIPA Buffer is compatible with this kit (Cat. No. 23225/23227).
Please refer to the link below for more information on this kit (Cat. No. 23225/23227).
https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0011430_Pierce_BCA_Protein_Asy_UG.pdf
23225 BCA™ Protein Assay Kit, sufficient reagents for 500 test tube or 5,000 microplate assays.
23227 BCA™ Protein Assay Kit, sufficient reagents for 250 test tube or 2,500 microplate assays.
Best.
I would use a BCA assay like advised by others, as the assay is unaffected by detergents and the majority of common buffer components (excluding reducing agents, which have to be avoided). I would especially look at the Pierce Rapid Gold BCA assay sold by Thermo, as it has a real advantage in being able to be analysed after only 5 minutes at room temperature rather than 30+ minutes at elevated temperatures like conventional BCA assays.
Yes, RIPA buffer is compatible with the Thermo Scientific Pierce Modified Lowry protein quantification assay.
- Badges
- Science method