Excellent Post-Conjugation Cleanup with High Recovery and Minimal Sample Loss

June 22, 2026

BlueskyReddit
MD Anderson Cancer Center - University of Texas
Cancer System Imaging (Diagnostic Imaging Division)
Lab Technician

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Company:

Zeba™ Spin Desalting Columns, 7K MWCO, 5ml

Product Name:

Thermo Fisher Scientific

Catalog Number:

89891

Zeba Spin Desalting Columns (7K MWCO, 5 mL) are my go-to for purifying antibody-dye conjugates post-conjugation. Clean, fast, and consistently reliable — these columns never disappoint.

Experimental Design and Results Summary

Application

Post-conjugation purification of antibody-dye conjugates, buffer exchange, and removal of unconjugated free dye or small molecules.

Starting Material

Crude antibody-dye conjugation reaction mix (~120 µL), diluted with 40 µL PBS prior to loading as per protocol recommendation. Antibody MW ~150 kDa, well above the 7K MWCO cutoff, ensuring full antibody retention while free dye passes through.

Tips

Add 40 µL of PBS to your ~120 µL sample before loading — as suggested by the protocol — to optimize flow and recovery Equilibrate the column with your desired storage buffer before loading Stick to the recommended sample volume — don't overload Spin at the exact recommended speed and time for best recovery Run a quick A280/dye absorbance check after elution to confirm purity and degree of labeling

Results Summary

Excellent free dye removal with high antibody recovery in a single spin. The 7K MWCO is ideal for antibodies (~150 kDa), retaining the conjugate completely while efficiently removing unconjugated dye. Conjugate comes out clean and ready for downstream use — no additional purification steps needed.

DOI or PMID #

N/A

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Summary

The Good

Fast, reliable, and reproducible. It removes unconjugated free dye efficiently while fully preserving antibody integrity at ~150 kDa.

The Bad

Single-use only, which adds up in cost for high-throughput conjugation workflows.

The Bottom Line

If you're doing antibody-dye conjugations, these columns are a must-have. Load your sample, follow the simple protocol, one spin and your conjugate is ready to go — it simply works.

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