Industry-leading recovery of PBMCs allows you to get the most out of your leukopaks
Gently isolate truly untouched Human Peripheral Blood Mononuclear Cells (PBMCs) directly from leukapheresis material in under an hour without the need for lysis or density gradient centrifugation. The Human PBMC Leukopak Isolation Kit was developed with BACS™ Microbubbles to isolate untouched leukopak PBMCs from leukapheresis material. Non-PBMCs are targeted and removed with antibodies recognizing CD66b, CD123, and CD235ab via negative selection. Isolated PBMCs are suitable for flow cytometry, molecular assays, activation and expansion, cell culture, and other functional studies.
The Human PBMC Leukopak Isolation Kit has the capacity to isolate PBMCs from 12.5 x 109 total cells (an average half-leukopak, red blood cells and white blood cells combined).
The Human PBMC Leukopak Isolation Kit, Research Scale, has the capacity to isolate PBMCs from 2.0 x 109 total cells (red blood cells and white blood cells combined).
Chemical-free processing that avoids exposing cells to lysis and density gradient reagents for uncovering more meaningful biology
Scalable: isolate PBMCs in tubes or directly in the leukopak bag
Healthier cells: less handling with only one wash step no matter the blood cell amount
How it Works: Leukopak samples contain a milieu of immune cells (i.e., white blood cells), plasma, platelets, and red blood cells. The Human PBMC Leukopak Isolation Kit uses a chemical-free, gentle negative selection method to enrich leukopak PBMCs by removing unwanted cell populations.
These unwanted cells are labeled with an optimized cocktail of biotinylated antibodies targeting CD66b, CD123, and CD235ab. BACS™ Streptavidin Microbubbles are mixed into the sample where they bind to the labeled cells. Due to their inherent buoyancy, the microbubbles quickly and gently float the non-PBMCs to the surface. Following centrifugation, the PBMCs are available for collection.
The Human PBMC Leukopak Isolation Kit minimizes handling steps and consistently delivers more isolated PBMCs. Resulting PBMCs from the Akadeum kit have the benefit of being untouched and not being exposed to chemical processes. Isolated leukopak PBMCs are suitable for flow cytometry, molecular assays, activation and expansion, cell culture, cell engineering or other functional studies.
Additional Information:
Storage: 4°C
Cell Type: PBMCs
Sample Species and Source: washed human leukopak apheresis material
Cell Separation Method: Negative Selection
Capacity:
Human PBMC Leukopak Isolation Kit: 12.5 x 109 total cells (enough to process an average half-leukopak)
Human PBMC Leukopak Isolation Kit, Research Scale: 2.0 x 109 total cells
Research Use Only
This Kit Comes With:
Human PBMC Leukopak Isolation Kit:
62.5 mL BACS™ Streptavidin Microbubbles
12.5 mL Human PBMC Leukopak Biotin-Antibody Cocktail
500 mL Separation Buffer (Ca2+ and Mg2+ free PBS containing 2 mM EDTA, 0.5% biotin-free BSA)
Human PBMC Leukopak Isolation Kit, Research Scale:
10 mL BACS™ Streptavidin Microbubbles
1 mL Human PBMC Leukopak Biotin-Antibody Cocktail
200 mL Separation Buffer (Ca2+ and Mg2+ free PBS containing 2 mM EDTA, 0.5% biotin-free BSA)
Scale your research from tube to bag. Just use more or less to suit your research needs.
Example of scale: 50 million cells in a tube, to 10 billion cells in bag.
For running the Human PBMC Leukopak Isolation Kit on the Alerion™, please contact Technical Support.
Data
Isolated PBMCs were labeled for CD45 and CD235a. The fluorescently labeled cells were analyzed by flow cytometry. Dead cells and debris were excluded from the analysis.
Resources
Protocol Sheet
Publications & References
FAQs
Research and Purification Products
1. Can I do back-to-back isolation to improve purity?
Yes. This can be specifically helpful when using the Dead Cell Removal Microbubble Kit or the Human Red Blood Cell Depletion Microbubbles.
2. Which cell types can be processed using Akadeum’s Dead Cell Removal Microbubble Kit?
Akadeum’s Dead Cell Removal Microbubble Kit utilizes Annexin V-coated microbubbles to selectively bind phosphatidylserine, a marker exposed on the surface of cells undergoing apoptosis. This technology enables efficient removal of apoptotic cells across various cell types where apoptosis is actively occurring.
3. Can the Dead Cell Removal Microbubble Kit be used if the sample is under 35% viability?
Although the initial cell viability is below the recommended threshold, successful outcomes have been achieved by performing the protocol twice on such samples.
General Questions
Are Akadeum microbubbles hollow silica spheres?
Yes, Akadeum’s microbubbles are hollow silica spheres. Silica is a material that has extensive precedence for use in therapeutics and diagnostics.
Are antibodies directly attached to the microbubbles?
Only specific kits, such as our Dead Cell Removal (DCR) and RBC microbubble kits, use antibody-coated microbubbles. Most Akadeum microbubble products are not coated with antibodies; instead, a biotinylated antibody cocktail is provided to use with the streptavidin-coated microbubbles.
What is buoyancy-activated cell sorting (BACS)?
BACS is Akadeum’s proprietary cell separation technology that uses buoyant microbubbles to isolate specific cell populations utilizing only the power of gravity.
What are microbubbles?
Akadeum’s microbubbles are hollow silica spheres that are on average 12 microns.
How big are microbubbles?
Microbubbles average 12 um in diameter. Learn more about microbubbles technology.
How many cells can a microbubble lift?
Microbubbles can carry as many as four cells.
What is the Separation Buffer?
Ca2+, Mg2+ free Dulbecco’s PBS containing 2mM EDTA, and 0.5% bovine serum albumin (BSA). BSA must be biotin free to not interfere with kit performance.
After sitting in the refrigerator, all the microbubbles have separated out of solution. Is there something wrong?
No, this is normal due to the buoyant properties of microbubbles. The solution will need to be mixed well before every use. If processing multiple tubes, mixing between tubes is necessary.
I don’t have an aspirator/vacuum available in my lab, can I still use Akadeum microbubbles?
Aspiration is the best and easiest way to remove the microbubble layer after centrifugation. Pipetting off the bubble layer is not very effective. Vacuum pumps are available from a variety of vendors.
How are Akadeum’s kits shipped?
Kits are shipped on ice packs with overnight delivery.
I have additional technical questions, who should I contact?
Please email all technical questions to techsupport@akadeum.com or check out our chatbot!
