Sample Preparation



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  1. Mammalian Tissue Protein Extraction Reagent AR0101
    $100.00
    • Western Blotting Reagents, Sample Preparation
    • 100 mL (100 assays for 0.1g of tissuе)
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    Boster’s Mammalian Tissue Protein Extraction Reagent is a ready-to-use Western blot related reagent solution used for efficient extraction of total soluble protein in nondenatured state from animal tissues. Learn More
  2. ISH Detection Kit (POD) MK1030
    $150.00
    • Western Blotting Reagents, Sample Preparation
    • 1 kit (sufficient for staining100 slides)
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    ISH Detection Kit Learn More
  3. Bovine Serum Albumin Standard AR0191
    $20.00
    • Western Blotting Reagents, Sample Preparation
    • 20mL
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    Bovine Serum Albumin Standard is a ready-to-use solution for determining protein concentration Learn More
  4. Broad Spectrum Protease Inhibitor Cocktail (EDTA free) (100x) AR1182-1
    $50.00
    • Western Blotting Reagents, Sample Preparation
    • 1ml
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    Boster’s Broad Spectrum Protease Inhibitor Cocktail (EDTA free) is a complex of various protease inhibitors, which has been tested for inhibiting proteases and esterase broadly.  Learn More
  5. Proteinase K AR0056
    $50.00
    • Western Blotting Reagents, Sample Preparation
    • 5ml
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    Boster’s Proteinase K is a Tris-HCl buffered concentrated enzyme stock solution that is to be used as assistant pre-treatment reagent in Western blot and IHC assay procedures. Learn More
  6. Broad Spectrum Phosphatase Inhibitor Cocktail AR1183
    $30.00
    • Western Blotting Reagents, Sample Preparation
    • 1 ml, 100X concentrated stock solution
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    Broad Spectrum Phosphatase Inhibitor Cocktail, For inhibiting Protein tyrosine phosphatase, acidic and alkaline phosphatase; Serine/threonine phosphatase, acidic and alkaline phosphatase; PP1and PP2A. Learn More
  7. Broad Spectrum Protease Inhibitor Cocktail (100x) AR1182
    $50.00
    • Western Blotting Reagents, Sample Preparation
    • 1ml
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    Broad Spectrum Protease Inhibitor Cocktail, For inhibiting proteases and esterase. Learn More
  8. RIPA Lysis Buffer AR0105
    $25.00
    • Western Blotting Reagents, Sample Preparation
    • 50ml
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    RIPA Lysis Buffer reagent is a complete cell lysis reagent popularly used for cultured mammalian cells. RIPA lysis buffer is highly compatible with immunoassays, protein purification procedures, immunoprecipitation, and western blotting. Learn More
  9. Phosphatase Inhibitor Cocktail III AR1121
    $15.00
    • Western Blotting Reagents, Sample Preparation
    • 80mg, enough for 100ml of samples
    • boster_tick.pngShips next business day
    Boster’s Phosphatase Inhibitor Cocktail III is a Western blot related that is to be added to cell lysis buffer to protect native phosphoproteins from dephosphorylation during proteins purification and sample preparation used in WB, Co-IP, and IP. Learn More
  10. Phosphatase Inhibitor Cocktail II AR1120
    $20.00
    • Western Blotting Reagents, Sample Preparation
    • 80mg, enough for 100ml of samples
    • boster_tick.pngShips next business day
    Boster’s Phosphatase Inhibitor Cocktail II is a Western blot related that is to be added to cell lysis buffer to protect native phosphoproteins from dephosphorylation during proteins purification and sample preparation used in WB, Co-IP and IP. Learn More

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Western Blotting Sample Preparation Products

western blot prep

Introduction

Western blotting among the most frequently employed techniques used to detect proteins based on antigen-antibody interactions. The first step of western blotting is sample preparation; your protein of interest must be extracted from the tissue or cell culture it’s expressed in, protected from degradation, and prepared to be run on a gel. Boster provides products for sample preparation, including specialized extraction kits, lysis buffers, and protease inhibitors.

Procedure

To extract a protein sample from a cell culture on solid medium, first rinse the culture with ice-cold PBS solution, then add ice-cold lysis buffer. Scrape adherent cells off the dish using a cold plastic scraper, or digest the cells with 0.05% trypsin. Once the cells are suspended, centrifuge the sample, draw off the supernatant fluid, and wash the pellet with ice cold PBS buffer. Re-suspend the cells in Boster mammalian cell protein extraction reagent and sonicate the solution for about 10-15 seconds, then lyse the cells with Boster RIPA lysis buffer and additional sonication. Centrifuge the sample to separate lipids and cellular debris from the soluble cell proteins, and aliquot the middle layer from the centrifuged sample into a fresh tube and store it at -20C.

The aliquots of cell lysis supernatant are ready for total protein quantification and gel electrophoresis.

For a detailed protocol of sample preparation starting with tissue samples, see our Western blot handbook.

Tips

  1. For best results, select the right type extraction kit for your sample
  2. If you observe weak signal, ghost bands, or inconsistent band sizes in your results, the source of error may be the protein extraction step. See our troubleshooting guides for more information
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