⚗️ Buffer Recipe Calculator

Precision is key. Standard recipes for SPR and BLI running buffers. Calculations use accurate molecular weights for standard lab salts (e.g., EDTA disodium dihydrate).

Running buffer = sample buffer. Always dilute your analyte into the same buffer used for the run — matched ionic strength, surfactant, and DMSO content are critical for clean kinetics.

Standard Biacore running buffer. Good for general protein interactions.

HBS-EP+

Target pH: 7.4
1000 mL
Final Volume
ReagentAmount
HEPES
MW: 238.30 g/mol
2.38 g
NaCl
MW: 58.44 g/mol
8.77 g
EDTA (Disodium)
MW: 372.24 g/mol
1.12 g
Surfactant P20 / Tween 20
From 10% Stock Solution (prepare: 10 g Tween 20 + water to 100 mL)
5.00 mL

Preparation Steps:

  1. Dissolve reagents in approx 800 mL of ultrapure water (80% of final vol).
  2. Adjust pH to 7.4 using NaOH.
  3. Cytiva Surfactant P20 is a polysorbate-20 formulation; generic Tween 20 (polysorbate 20) is an acceptable substitute.
  4. Add water to final volume of 1000 mL.
  5. Filter through 0.22 µm membrane. Degas if using for SPR.

Common Regeneration Buffers

💡 Key principle: For acid regeneration (glycine), pH dominates over buffer concentration. For chaotropes (guanidine-HCl) and detergents (SDS), concentration is the primary variable — 6 M GuHCl is far harsher than 1 M. Start mild, increase harshness only if needed.

BufferRecipeHarshness
Glycine-HCl pH 2.010 mM glycine + HCl to pH 2.0
Glycine-HCl pH 1.510 mM glycine + HCl to pH 1.5
NaOH 10 mM10 mM NaOH in water
NaOH 50 mM50 mM NaOH in water
NaCl 1–2 M1–2 M NaCl in running buffer
MgCl₂ 1–4 M1–4 M MgCl₂ in water (antibody / electrostatic capture surfaces)
6M Guanidine-HCl6 M guanidine hydrochloride in water
0.1% SDS0.1% (w/v) sodium dodecyl sulfate in water

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