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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.