Applicazione
Proteomics buffers
Buffers, reducing agents, detergents and chaotropes for lysis, reduction, denaturation and resolution.
Proteomics workflows rely on buffers (Tris, HEPES, phosphate), reducing agents (DTT, TCEP), detergents (SDS, Triton X-100) and chaotropes (urea, guanidine HCl) to lyse cells, keep proteins soluble, break disulfides, and resolve them on a gel or column. Molekula supplies these reagents in research and bulk quantities with ISO 9001:2015 traceability and SDS in five languages.
Cosa è incluso in questo flusso di lavoro
- Lysis & extraction
- Tris-HCl, HEPES, sodium phosphate, NaCl, EDTA, protease inhibitor cocktails.
- Reduction
- DTT (dithiothreitol) and TCEP cleanly cleave protein disulfide bonds. TCEP is preferred for mass spec because it does not react with iodoacetamide.
- Denaturation
- SDS for full denaturation; urea and guanidine HCl when native fold needs to be unfolded reversibly.
- Detergents
- Triton X-100, NP-40, CHAPS, sodium deoxycholate, n-dodecyl β-D-maltoside.
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FAQ
Which reducing agent should I use, DTT or TCEP?
TCEP is more stable at neutral and slightly acidic pH and does not interfere with cysteine alkylation in mass spec workflows. DTT is cheaper and works well for short reactions at pH 7.5–9. Use TCEP for MS sample prep, DTT for SDS-PAGE sample buffer.
What buffer pH range works for most proteins?
Most biochemistry runs at pH 7.4–8.0 using Tris or HEPES. Pick a buffer whose pKa is within ±1 unit of your target pH (Tris pKa 8.1, HEPES pKa 7.5, phosphate pKa 7.2).
How do I prevent oxidation of cysteines?
Keep 1–10 mM DTT or 1 mM TCEP in your buffer, work at lower pH if possible, and degas with nitrogen for sensitive proteins.
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