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📏 Protein Science · Method

BCA Protein Quantification

Colorimetric protein quantification
🎯 Difficulty ★☆☆ Easy Duration About 1 h 🎯 Use Measure total protein concentration for loading normalization, enzyme assays and interactions
📖 Principle
Peptide bonds reduce Cu²⁺ to Cu⁺ in alkaline conditions; BCA chelates forming purple color proportional to concentration
Below are 4 steps. Open each to see how to do it and add products to your shared list.

📋 Workflow

Full workflow, products add-to-cart directly
1

Prepare standard curve

15 min
Dilute BSA standard into a gradient to build a concentration-absorbance calibration curve
💡 Tip Standards quality determines accuracy
⚠️ Caution Match sample buffer system
🧪 Materials for this step (click to shop by spec/brand)
2

Prepare BCA working reagent

10 min
Mix reagent A and B at ratio, store at RT in dark
💡 Tip Prepare fresh to avoid degradation
⚠️ Caution Protect from light for even color
🧪 Materials for this step (click to shop by spec/brand)
3

Dispense and incubate

30 min
Mix samples with working reagent, incubate 30 min at 37°C for color development
💡 Tip Dilute concentrated samples and factor in dilution
⚠️ Caution Reducing agents interfere with color development
🧪 Materials for this step (click to shop by spec/brand)
4

Read and calculate

10 min
Read at 562 nm on plate reader, calculate concentrations from the standard curve
💡 Tip Samples fall in linear range
⚠️ Caution Subtract blank background
🧪 Materials for this step (click to shop by spec/brand)

⚠️ 5 Common Beginner Mistakes

  • Reducing agents (DTT) interfere
  • High detergent concentration interferes
  • Buffer mismatch between standards and samples
  • Sample beyond linear range
  • Light-degraded reagent gives uneven color

❓ FAQ

+BCA vs Bradford?
BCA tolerates most detergents and is robust; Bradford is faster but detergent-sensitive.
+How to measure with reducing agents?
Switch to Bradford (reducing-agent compatible) or remove/dilute the reducing agent.
+Precision across concentration range?
Dilute high, load more for low, stay in mid-linear range for accuracy.

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Data authenticity: Product categories from JW Supply Chain standard taxonomy; method→step→product connected. Last updated: 2026-08-23 · v3.0