A Ponceau S Staining-Based Dot Blot Assay for Rapid Protein Quantification of Biological Samples
Despite the availability of a wide range of commercial kits, protein quantification is often unreliable, especially for tissue-derived samples, leading to uneven loading in subsequent experiments. Here we show that the widely used Bicinchoninic Acid (BCA) assay tends to underestimate protein concent...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-01-01
|
| Series: | Gels |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2310-2861/8/1/43 |
| _version_ | 1797493950371594240 |
|---|---|
| author | Dario Lucas Helbing Leopold Böhm Nova Oraha Leonie Karoline Stabenow Yan Cui |
| author_facet | Dario Lucas Helbing Leopold Böhm Nova Oraha Leonie Karoline Stabenow Yan Cui |
| author_sort | Dario Lucas Helbing |
| collection | DOAJ |
| description | Despite the availability of a wide range of commercial kits, protein quantification is often unreliable, especially for tissue-derived samples, leading to uneven loading in subsequent experiments. Here we show that the widely used Bicinchoninic Acid (BCA) assay tends to underestimate protein concentrations of tissue samples. We present a Ponceau S staining-based dot-blot assay as an alternative for protein quantification. This method is simple, rapid, more reliable than the BCA assay, compatible with biological samples lysed in RIPA or 2x SDS gel-loading buffer, and also inexpensive. |
| first_indexed | 2024-03-10T01:27:20Z |
| format | Article |
| id | doaj.art-f998c2f9228e44f5acd5a6518d34fa69 |
| institution | Directory Open Access Journal |
| issn | 2310-2861 |
| language | English |
| last_indexed | 2024-03-10T01:27:20Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Gels |
| spelling | doaj.art-f998c2f9228e44f5acd5a6518d34fa692023-11-23T13:50:22ZengMDPI AGGels2310-28612022-01-01814310.3390/gels8010043A Ponceau S Staining-Based Dot Blot Assay for Rapid Protein Quantification of Biological SamplesDario Lucas Helbing0Leopold Böhm1Nova Oraha2Leonie Karoline Stabenow3Yan Cui4Leibniz Institute on Aging—Fritz Lipmann Institute, 07745 Jena, GermanyLeibniz Institute on Aging—Fritz Lipmann Institute, 07745 Jena, GermanyLeibniz Institute on Aging—Fritz Lipmann Institute, 07745 Jena, GermanyFaculty of Medicine, Friedrich-Schiller-University Jena, 07743 Jena, GermanyLeibniz Institute on Aging—Fritz Lipmann Institute, 07745 Jena, GermanyDespite the availability of a wide range of commercial kits, protein quantification is often unreliable, especially for tissue-derived samples, leading to uneven loading in subsequent experiments. Here we show that the widely used Bicinchoninic Acid (BCA) assay tends to underestimate protein concentrations of tissue samples. We present a Ponceau S staining-based dot-blot assay as an alternative for protein quantification. This method is simple, rapid, more reliable than the BCA assay, compatible with biological samples lysed in RIPA or 2x SDS gel-loading buffer, and also inexpensive.https://www.mdpi.com/2310-2861/8/1/43protein quantificationgel electrophoresisWestern blotdot blot |
| spellingShingle | Dario Lucas Helbing Leopold Böhm Nova Oraha Leonie Karoline Stabenow Yan Cui A Ponceau S Staining-Based Dot Blot Assay for Rapid Protein Quantification of Biological Samples Gels protein quantification gel electrophoresis Western blot dot blot |
| title | A Ponceau S Staining-Based Dot Blot Assay for Rapid Protein Quantification of Biological Samples |
| title_full | A Ponceau S Staining-Based Dot Blot Assay for Rapid Protein Quantification of Biological Samples |
| title_fullStr | A Ponceau S Staining-Based Dot Blot Assay for Rapid Protein Quantification of Biological Samples |
| title_full_unstemmed | A Ponceau S Staining-Based Dot Blot Assay for Rapid Protein Quantification of Biological Samples |
| title_short | A Ponceau S Staining-Based Dot Blot Assay for Rapid Protein Quantification of Biological Samples |
| title_sort | ponceau s staining based dot blot assay for rapid protein quantification of biological samples |
| topic | protein quantification gel electrophoresis Western blot dot blot |
| url | https://www.mdpi.com/2310-2861/8/1/43 |
| work_keys_str_mv | AT dariolucashelbing aponceausstainingbaseddotblotassayforrapidproteinquantificationofbiologicalsamples AT leopoldbohm aponceausstainingbaseddotblotassayforrapidproteinquantificationofbiologicalsamples AT novaoraha aponceausstainingbaseddotblotassayforrapidproteinquantificationofbiologicalsamples AT leoniekarolinestabenow aponceausstainingbaseddotblotassayforrapidproteinquantificationofbiologicalsamples AT yancui aponceausstainingbaseddotblotassayforrapidproteinquantificationofbiologicalsamples AT dariolucashelbing ponceausstainingbaseddotblotassayforrapidproteinquantificationofbiologicalsamples AT leopoldbohm ponceausstainingbaseddotblotassayforrapidproteinquantificationofbiologicalsamples AT novaoraha ponceausstainingbaseddotblotassayforrapidproteinquantificationofbiologicalsamples AT leoniekarolinestabenow ponceausstainingbaseddotblotassayforrapidproteinquantificationofbiologicalsamples AT yancui ponceausstainingbaseddotblotassayforrapidproteinquantificationofbiologicalsamples |