Introduction
Antibodies are sensitive biological reagents, and their performance can be significantly affected by how they’re handled and stored. Even the most well-validated antibody can yield poor results if it has been degraded, contaminated, or repeatedly thawed and refrozen.
Whether you’re working with monoclonal, polyclonal, or recombinant antibodies, adopting proper storage and handling practices is essential to ensure long-term stability, experimental reproducibility, and data integrity. This guide walks through best practices for storage conditions, buffer compatibility, freeze-thaw handling, and stability-enhancing additives.
1. Understanding Antibody Stability
Antibodies, being proteins, are subject to:
- Denaturation due to heat or pH changes
- Aggregation during freeze-thaw cycles
- Microbial contamination when stored improperly
- Proteolytic degradation in the absence of stabilizers
The stability of an antibody depends on:
- Isotype and species origin
- Purification method
- Storage buffer composition
- Storage temperature
2. Recommended Storage Temperatures
| Storage Duration | Temperature | Notes |
| Short-term (≤1 month) | 2–8°C (refrigerator) | Ideal for frequent use; avoid freeze-thaw cycles |
| Medium-term (1–12 months) | –20°C | Aliquot to avoid freeze-thaw; use glycerol if needed |
| Long-term (>1 year) | –80°C | For extended storage, especially for rare clones or custom antibodies |
Important Notes:
- Avoid frost-free freezers which undergo temperature cycling
- Use dedicated storage boxes to minimize light and disturbance
- Always consult the manufacturer’s datasheet before deviating
3. Should You Freeze Antibodies?
Freezing is generally safe for most purified antibodies, but:
- Repeated freeze–thaw cycles cause aggregation and activity loss
- Conjugated antibodies (e.g., fluorophore-, enzyme-, or biotin-labeled) may degrade faster when frozen
- Crude antiserum or ascites fluid should not be frozen unless glycerol is added
Best Practice:
Aliquot antibodies into small volumes (10–50 µL) before freezing so you only thaw what you need.
4. Antibody Storage Buffers: What to Know
Common Buffer Components:
| Component | Purpose | Note |
| PBS / Tris | Maintains pH and ionic strength | Most standard antibodies use these buffers |
| BSA or gelatin | Stabilizer and protein filler | Avoid in conjugation reactions |
| Glycerol (50%) | Cryoprotectant for –20°C storage | Prevents freezing; allows use directly from freezer |
| Sodium azide (0.02–0.05%) | Antimicrobial | Toxic to live cells; avoid in cell-based assays |
| Proclin / Thimerosal | Preservatives | Less common; check for downstream compatibility |
Avoid:
- Metal ions or chelating agents (can inactivate HRP)
- Detergents or urea unless required for solubilization
5. Special Handling for Conjugated Antibodies
Conjugated antibodies (e.g., HRP, FITC, PE, Alexa Fluor) require more careful handling:
- Store in the dark to prevent photobleaching
- Use amber tubes or wrap in foil
- Avoid freeze–thaw altogether if possible; store at 4°C with azide + BSA
- Do not spin-concentrate fluorophore-labeled antibodies—they’re prone to sticking
6. Lyophilized Antibodies
Some antibodies are shipped lyophilized (freeze-dried) for improved stability at room temperature. Once reconstituted:
- Use the recommended reconstitution buffer (usually sterile water or PBS)
- Filter sterilize if you plan to store long term
- Store as per guidance (often –20°C or –80°C with glycerol)
7. Best Practices in Antibody Handling
- Always use sterile technique when accessing stock vials
- Avoid frothing or vortexing—gentle mixing is best
- Label aliquots clearly with concentration, clone, date, and freeze-thaw count
- Use low-binding tubes to minimize protein loss
8. Stability Enhancers: Should You Use Them?
Commercial antibodies often include stabilizers like:
- BSA
- Glycerol
- Sucrose
- Trehalose
If formulating your own antibody:
- Consider adding 0.1–1% BSA and 50% glycerol for –20°C stability
- Avoid preservatives if the antibody will be used in live-cell applications
Frequently Asked Questions
Can I store antibodies at room temperature?
Only lyophilized antibodies designed for ambient shipping. Reconstituted or liquid antibodies should be stored at 2–8°C or below.
Can I autoclave or filter antibodies?
Do not autoclave. You may 0.22 µm filter them under sterile conditions if needed, but some antibody loss may occur.
How do I know if my antibody has gone bad?
Signs include:
- Precipitation or cloudiness
- Poor signal in previously optimized assays
- Increased background or non-specific bands
Conclusion
Proper storage and handling of antibodies is essential to maintain activity, reduce background, and preserve reproducibility. By understanding buffer composition, conjugate sensitivity, and the risks of freeze-thaw damage, you can extend the life of your reagents and protect the integrity of your experiments.
At KinesisDx, all antibodies are shipped with optimized buffers and detailed handling recommendations. We help researchers protect their investment—experiment after experiment.
Works Cited
- Harlow, Ed, and David Lane. Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory Press, 1988.
- Greenfield, Elizabeth A. Antibodies: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, 2014.
- Abcam. “Best Practices for Antibody Storage.” Antibody Handling Guide, 2025, https://www.abcam.com.
- Bio-Rad. “Storage Recommendations for Antibodies and Conjugates.” 2025, https://www.bio-rad-antibodies.com.
Wikimedia Commons. “Protein Storage Best Practices.” https://commons.wikimedia.org/wiki/File:Protein_storage_best_practices.png
