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Published TCIMAIL newest issue No.198
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The first step in protein purification, lysing tissue or cells to extract protein, is crucial for properties and yield of the obtained protein. We offer detergent-based extraction buffers optimized for a wide range of target tissues, from cultured cells to nervous tissue and microorganisms.
Products
Advantages
- The target protein can be obtained easily by suspending tissue or cells and centrifuging.
- Extracted protein can be used directly in downstream analysis such as Western blotting.
- Product lineup : suitable for a wide range of subjects, from cultured cells to nervous tissue and microorganisms.
Application using Nervous Tissue Protein Extraction Buffer (Product No. B6279)
- Wash mouse brain twice with PBS and weigh samples.
- For each gram of sample, add 10 mL of Nervous Tissue Protein Extraction Buffer (Product No. B6279) and homogenize the sample.
- Incubate on ice for 10 minutes.
- Centrifuge the sample at 10,000 x g for 10 minutes at 4 °C.
- Analyze the supernatants directly by Western blotting.
Figure 1. Western blot image of proteins extracted from mouse brain
Synaptophysin (38 kDa) can be extracted from mouse brain efficiently.
Application using RIPA Buffer (Product No. R0246)
- Wash the cultured mammalian cells twice with PBS.
- Resuspend cells in RIPA Buffer (Product No. R0246). Use 1 mL buffer per 0.5 - 5.0 × 107 cells.
- Incubate on ice for 15 minutes.
- Centrifuge the sample at 10,000 x g for 10 minutes at 4 °C.
- Analyze the supernatants directly by Western blotting.
Figure 2. Western blot image of proteins extracted from mammalian cells
β-Actin (42 kDa) can be extracted from mammalian cells efficiently.
Movie of Protein Extraction Protocol for Mammalian Cells using RIPA Buffer (Product No. R0246)
Application using E.coli / Yeast Protein Extraction Buffer (Product No. Y0021)
- Centrifuge E. coli at 5,000 x g and yeast at 3,000 x g for 10 minutes. Remove as much medium as possible from pellet.
- Resuspend pellet in E.coli / Yeast Protein Extraction Buffer (Product No. Y0021). Use 2 - 4 mL buffer per gram pellet.
- Mix at room temperature for 10 minutes.
- Centrifuge E. coli at 15,000 x g and yeast at 14,000 x g for 10 minutes.
- Analyze the supernatants directly by Western blotting.
Figure 3. Western blot image of proteins extracted from E. coli and S. cerevisiae
RecA in E. coli (left, 38 kDa) and Rad51 in S. cerevisiae (right, 43 kDa) can be extracted efficiently.