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    Protocols
    Bioss is dedicated to helping you achieve exceptional results. Our detailed listings for reagnets and buffers for your application can be found below.



     



    Reagents and Bufferspdf iconDOWNLOAD A PDF

    Phosphate-Buffered Saline (PBS)(10x)
    • 80g of NaCl
    • 2.0g of KCl
    • 14.4g of Na2HPO4
    • 2.4g of KH2PO4
    • Mix 800mL ultra-pure water and adjust pH to 7.6 with pure HCl. Top up with ultra-pure water to 1L.
    Phosphate-Buffered Saline w/ Tween20 (PBST)
    • For 1L: 100mL of PBS 10x + 890mL ultra-pure water + 10 mL Tween 20
    Tris-Buffered Saline (TBS) (10X)
    • 24.23g Trizma HCl
    • 80.06g NaCl
    • Mix in 800mL ultra-pure water and adjust pH to 7.6 with pure HCl. Top up with ultra-pure water to 1L.
    Tris-Buffered Saline w/ Tween20 (TBST)
    • For 1L: 100mL of TBS 10x + 890mL ultra-pure water + 10 mL Tween 20


    Western Blotting Buffers

    RIPA Buffer
    • 150mM NaCl
    • 1.0% NP-40 or 0.1% Triton X-100
    • 0.5% sodium deoxycholate
    • 0.1% SDS (sodium dodecyl sulphate)
    • 50mM Tris-HCl pH8.0
    • Protease Inhibitors
    Tris-HCl buffer
    • 20mM Tris-HCl pH7.5
    • Protease Inhibitors
    Denaturing lysis buffer
    • 1% SDS
    • 5mM EDTA
    • Immediately before use add:
      • 10mM dithiothreitol or beta-mercaptoethanol
      • Protease inhibitors
      • 15U/mL DNase1
    Laemmli 2X buffer/loading buffer
    • 4%SDS
    • 10% 2-mercaptoethanol
    • 20% glycerol
    • 0.004% bromophenol blue
    • 0.125M Tris-HCl
    • Check the pH and adjust pH to 6.8.
    Running buffer
    • 25mM Tris base
    • 190mM glycine
    • 0.1% SDS
    • Check the pH, which should be about pH 8.3. Adjust if necessary.
    Transfer buffer (wet)
    • 25mM Tris base
    • 190mM glycine
    • 0.1% SDS
    • The pH should be about pH8.3. Adjust if necessary.
    Transfer buffer (semi-dry)
    • 48mM Tris
    • 39mM glycine
    • 20% methanol
    • 0.04% SDS
    Blocking buffer
    • 5% milk or BSA(bovine serum albumin)
    • Add to TBST buffer. Mix well and filter.

     



    Immunohistochemistry/Immunocytochemistry Buffers

    Formalin solution (10%)
    • 3.7-4% Formaldehyde (37-40%)
    • 33mM NaH2PO4
    • 46mM Na2HPO4
    Paraformaldehyde (4%)
    • 8% paraformaldehyde
    0.2M Phosphate Buffer(PB) pH7.4
    • 53mM NaH2PO4
    • 154mM Na2HPO4
    • Heat 8%PFA solution at 60C while stirring. Once the solution reaches 60C and the PFA dissolves, add 500mL of 0.2M phosphate buffer, to bring the solution to 4%PFA in 0.1M phosphate. Carefully add 1N NaOH until the solution is clear. Cool the solution and filter.
    Sodium Citrate Buffer pH6.0
    • 10mM sodium citrate
    • 0.05% Tween20
    • Mix to dissolve sodium citrate and adjust pH to 6.0 with 1N HCl.
    • Add Tween20 and mix well.
    • Store at room temperature for 3 months or at 4C for longer storage.

     



    ELISA Buffers

    Bicarbonate/carbonate coating buffer (1oomM) pH9.6
    • Antigen or antibody should be diluted in coating buffer to immobilize them to the wells:
    • 29mM Na2CO3
    • 71mM NaHCO3
    Blocking solution
    • Commonly used blocking agents are 1% BSA, serum, or non-fat dry milk in PBS.
    Wash solution
    • Usually PBS or Tris-buffered saline(pH7.4) with detergent such as 0.05% (v/v) Tween 20 (TBST).

     



    Flow Cytometry

    FACS buffer/antibody dilution buffer
    • 10% FCS
    • 1% sodium azide in PBS
    Permeabilization
    • 0.1-1% Triton X-100/NP-40 in PBS
    Fixative
    • 0.01-0.1% paraformaldehyde in PBS.