Choosing the Right Buffer for Cell Separation
When isolating cells using ·¡²¹²õ²â³§±ð±èâ„¢ or ¸é´Ç²ú´Ç³§±ð±èâ„¢ cell separation systems, the recommended buffer is 2% FBS in Phosphate Buffered Saline w/ 1 mM EDTA
When isolating cells using ·¡²¹²õ²â³§±ð±èâ„¢ or ¸é´Ç²ú´Ç³§±ð±èâ„¢ cell separation systems, using the right buffer is critical for achieving consistent, high-quality results. The recommended buffer is typically 2% fetal bovine serum (FBS) in phosphate-buffered saline (PBS; without Ca++ and Mg++) with 1 mM EDTA. For convenience and consistency, this buffer is available ready-made from º£½ÇÆƽâ°æ Technologies (·¡²¹²õ²â³§±ð±èâ„¢ Buffer, 1 L, Catalog #20144 or ¸é´Ç²ú´Ç³§±ð±èâ„¢ Buffer, 250 mL, Catalog #20104).
The following alternative base solutions can also be used:
- Dulbecco’s PBS (D-PBS; Without Ca++ and Mg++; 500mL, Catalog #37350 (1X) and Catalog #37354 (10X))
- Hank’s Balanced Salt Solution (HBSS), Modified (Without Ca++ and Mg++; 500 mL, Catalog #37250) may also be used as an alternative, though PBS or D-PBS remains the preferred choice
For Diagnostic Workflows
The ·¡²¹²õ²â³§±ð±èâ„¢ Buffer (IVD; 1 L, Catalog #100-0780) is intended for use with ·¡²¹²õ²â³§±ð±èâ„¢ cell isolation kits that are available as in vitro diagnostic (IVD) medical devices in specific regions.*
Buffer Options for Your Workflow
Different workflows may require different buffer compositions, including standard (FBS-based), serum-free, diagnostic, and custom buffer formulations. Select from the options optimized for ·¡²¹²õ²â³§±ð±èâ„¢ immunomagnetic cell separation and ¸é´Ç²ú´Ç³§±ð±èâ„¢ automated cell separation workflows below:
Standard Workflows
·¡²¹²õ²â³§±ð±èâ„¢ Buffer
Optimized for ·¡²¹²õ²â³§±ð±èâ„¢ cell separation workflows, this ready-to-use buffer formulation supports consistent performance.
Automated Workflows
¸é´Ç²ú´Ç³§±ð±èâ„¢ Buffer
Designed for use with ¸é´Ç²ú´Ç³§±ð±èâ„¢ automated systems, this buffer supports reproducible, walk-away cell separation.
Serum-Free Option
¸é´Ç²ú´Ç³§±ð±èâ„¢ Buffer 2
This serum-free formulation containing 0.5% BSA is suitable for workflows where serum components may interfere.
Diagnostic Workflows
·¡²¹²õ²â³§±ð±èâ„¢ Buffer (IVD)
Intended for use with ·¡²¹²õ²â³§±ð±èâ„¢ cell isolation kits available as IVD medical devices, this buffer supports washing of cells or dilution during enrichment of target cells for downstream diagnostic assays.*
Buffer Base Alternative
D-PBS (Without Ca++ and Mg++)
This phosphate-buffered saline without Ca++ and Mg++ can be used as a recommended base solution for custom cell separation buffer formulations.
Buffer Base Alternative
HBSS, Modified (Without Ca++ and Mg++)
This balanced salt solution without Ca++ and Mg++ can be used as a PBS substitute in buffer formulations.
Want to Avoid Serum?
FBS has an important role in maintaining cell quality during separation: it reduces cell aggregation and clumping, minimizes nonspecific binding to plasticware, and provides nutrients and growth factors to support cell health during handling. If you prefer to avoid serum, an alternative buffer can be used consisting of PBS (without Ca++ and Mg++) supplemented with 0.5% bovine serum albumin (BSA) or human serum albumin (HSA) and 2 mM EDTA. BSA and HSA are purified serum proteins that provide a more defined alternative to whole serum. We provide a ready-to-use serum-free option containing 0.5% BSA (¸é´Ç²ú´Ç³§±ð±èâ„¢ Buffer 2, 250 mL, Catalog #20164).
Why Include EDTA?
EDTA is an essential component of the cell separation buffer because it chelates divalent cations (Ca++ and Mg++). By binding these ions, EDTA minimizes unwanted cell-to-cell and cell-to-particle adhesion, resulting in cleaner separations and improved cell separation efficiency.
Use of Alternative Media
The use of alternative media (e.g. RPMI, DMEM/F12, or DMEM) is not recommended, as these have not been evaluated for cell separation applications.
Cell separation kits from º£½ÇÆƽâ°æ Technologies have been validated exclusively with the recommended buffers listed above. In addition:
- Most media (e.g. DMEM) contain Ca++, which can negatively affect cell separation performance
- Many media rely on CO2-dependent buffering systems and require a controlled environment with 5-10% CO2. Outside of a CO2 incubator, pH fluctuations may occur, which can further compromise cell separation
Not sure which buffer is right for your application? Our Product & Scientific Support team can help you optimize your workflow, at techsupport@stemcell.com.
Related Resources
Choosing a Cell Separation Method that Meets Your Research Needs
Learn more about the cell separation methods outlined above to choose the best method for your application.
On-Demand Human Immune Cell Isolation Course
Learn how to effectively process human samples and isolate highly pure target cells with ·¡²¹²õ²â³§±ð±èâ„¢ in this free self-paced course.
¸é´Ç²ú´Ç³§±ð±èâ„¢ Automated Cell Separation Instruments
Learn how to automate your cell processing and isolations using the ¸é´Ç²ú´Ç³§±ð±èâ„¢-S and ¸é´Ç²ú´Ç³§±ð±èâ„¢-16 cell isolation instruments.
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