Akadeum’s Human CD4+ T Cell Isolation Kit delivers a highly enriched population of healthy, viable CD4+ T cells using a protocol that is fast, easy, and exceptionally gentle on delicate immune cells.
This kit is designed to process 10 samples, up to 100 million cells per sample
Fast & easy workflow enriches human peripheral blood mononuclear cells (PBMCs) for CD4+ T cells in 30 minutes
No magnet, no column, no harsh chemicals
Microbubble enrichment maintains cell health and physiology
Akadeum’s Human CD4+ T Cell Isolation Kit delivers a high-purity population of human CD4+ T cells that are ready for downstream use. Use of these microbubbles to enrich human PBMC samples for CD4+ T cells prior to flow sorting can greatly reduce final sort times, helping to isolate more cells for better science. Akadeum’s microbubble workflow is exceptionally gentle on cells, helping to maintain the health and physiology of immune cells during enrichment.
Mechanism of Action:Akadeum’s Human CD4+ T Cell Isolation Kit uses a negative selection method to remove unwanted cell populations from the sample, effectively enriching for the T cells of interest while maintaining cellular physiology. The undesired (non-CD4+ T) cells are labeled with Akadeum’s optimized cocktail of biotinylated antibodies targeting CD8, CD11b, CD14, CD16, CD19, CD36, CD56, CD123, CD235a, and TCR-γδ. Streptavidin-coated microbubbles are then mixed into the sample, binding to the labeled cells and floating them to the top of the sample container for removal.
How it Works: First, Akadeum’s biotinylated antibody cocktail is added to the sample to label the contaminating cells. Next, the microbubbles are mixed into the sample where the seek out and bind to the unwanted cells. Once bound, they quickly and gently float the undesired cells to the surface. An optional centrifugation step pellets the CD4+ T cells, and the microbubbles, microbubble-bound cells, and supernatant are easily aspirated away leaving the untouched and enriched CD4+ T cells happy, healthy, and ready for downstream use.
Additional Information:
Storage: 4°C
Cell Type: Naïve / resting CD4+ T cells
Sample Species and Source: Fresh Human PBMCs
Cell Separation Method: Negative Selection
Capacity: 1 billion cells
This kit comes with:
Akadeum’s BACS™ Streptavidin Microbubbles
Akadeum’s Human CD4+ T Cell Antibody Cocktail
Separation Buffer (Ca2+ and Mg2+ free PBS containing 2 mM EDTA, 0.5% biotin-free BSA)
5mL Eppendorf Tubes
For best results, the following supplies are recommended:
Low retention 1mL pipet tips
Centrifuge with swinging bucket rotor
Vacuum aspirator
Key Features
Akadeum’s Human CD4+ T Cell Isolation Kit delivers a highly-enriched population of healthy, viable CD4+ T cells using our fast, easy, and gentle microbubble protocol. Ideal for enrichment of rare and delicate immune cells, Akadeum’s novel Buoyancy Activated Cell Separation (BACS™) technology retains precious cells during the enrichment process. Cell isolation occurs directly in the sample container, eliminating the need to expose rare and delicate cells to harsh chemicals or harmful shear forces. Because no additional equipment is required, cell loss is avoided and the health and physiology of Human CD4+ T cells is maintained. The end result is a highly-pure population of CD4+ T cells that are happy, healthy, and ready for downstream use.
Elevated Solutions for Sample Preparation:
Retain precious, rare cells and avoid cell loss
No magnet, no column, no additional equipment
Fast and easy workflow takes only 30 minutes
Highly enriched, viable human CD4+ T cells
Elegant in Simplicity, Powerful in Application
Akadeum’s microbubble technology harnesses the power of buoyancy in a fast, easy and gentle workflow without the need for additional equipment. This unique, gentle approach maintains cell health and physiology.
No magnet. No column. Happy cells.
We’ve taken technology that traditionally requires additional equipment to accomplish targeted cell separation and put it into a single container where it’s self-separating. No harmful shear forces, no exposure to external forces like magnetic gradients. Stop losing your precious cells to harsh, outdated processing techniques.
Experience the benefits of a microbubble workflow. Akadeum’s Microbubbles Are:
Magnet-free, protecting your cells from harsh magnetic forces and eliminating the need for additional (and expensive!) equipment
Column-free, using microbubbles directly in the sample vial to separate cells allows for quick and easy isolation of any sample, any volume, anywhere
Gentle on cells, harnessing the targeted efficiency of affinity molecules coupled with the power of buoyancy to gently separate via flotation
Reliable and consistent, delivering fast and easy results that can be scaled for larger volumes with workflows that are compatible with nearly any downstream processing
Looking for Answers?
At Akadeum Life Sciences, our team is committed to your success. We thrive on problem-solving and are dedicated to helping the researchers we work with see exceptional results. If you’re interested in leveraging the microbubble platform to overcome existing hurdles in your sample preparation processes, we want to hear from you! You can speak directly to our team of scientists who are available to help you with an optimized workflow that takes advantage of our microbubble technology and solves long-standing headaches in sample preparation. Schedule a call today, we’d love to start a conversation!
Data
Isolation of human CD4+ T cells from PBMCs was performed using Akadeum Life Sciences Human CD4+ T Cell Isolation Kit (#13210-130). Cells were labeled with CD4-PerCP/Cy5.5 and CD8-PE. Dead cells and debris were excluded from analysis.
Workflow in Use:
Regulatory T cells (Tregs) represent a rare and important cell population in the immune system. A highly specialized subpopulation of T cells, Tregs account for only 4-10% of the total CD4+ T cell population and constitute merely 1-5% of all peripheral blood mononuclear cells (PBMCs), making them a rare target of interest that can be challenging to isolate in a large enough population for meaningful downstream analysis. Because Tregs express CD4, a sample preparation step using Akadeum’s CD4+ T Cell Microbubbles allows for fast, easy, and exceptionally gentle enrichment prior to cell sorting.
Using Akadeum’s Human CD4+ T Cell Kit ahead of flow sorting can provide a highly-enriched population of CD4+ T cells from PBMC samples using a rapid, gentle, and efficient protocol that reduces final sort times 15-fold while improving cell viability and lowering costs.
Illustrative CD4+ T Cell Enrichment with Human CD4+ T Cell Isolation Kit from Akadeum Life Sciences. Isolation of human CD4+ T cells from PBMCs was performed, cells were labeled as indicated, and analyzed on a CytoFLEX flow cytometer. A.) Flow plots showing an example before and after CD4+ T cell enrichment. B.) Before and after CD4+ T cell enrichment results from one-day old PBMCs (n=7). Dead cells and debris were excluded from all analysis.
Using microbubbles and microfluidic flow cytometry to enrich Treg cells created a sample in which the CD4+ T cell population made up 95-98% of cells within the lymphocyte gage and Tregs within the lymphocyte population went from an average of 1% to 85-94%.
Sorting Enriched Sample with WOLF® Cell Sorter Yields High Purity Treg Population. A.) Percentage of CD4+ T prior to enrichment (green bars) and after microbubble enrichment sorting (blue bars). n=1-2 per donor. B.) Percentage of Tregs prior to enrichment (green bars) and after microbubble enrichment sorting (blue bars). n=1-2 per donor.
Using Akadeum’s quick and easy CD4+ T cell enrichment microbubbles prior to Treg isolation via cell sorting led to a significant reduction in the overall time needed to obtain a sufficient number of pure, viable Treg cells for downstream applications like single-cell sequencing. The microbubbles combined with NanoCellect’s WOLF® sorter maintained exceptional purity, reduced enrichment times 15-fold, and increased sorting speed without introducing additional shear stress.
How to Use Akadeum’s Human CD4+ T Cell Isolation Microbubbles:
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!
