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ReadiPrep™ CD4+ Cell Isolation Kit *Optimized for processing 30 mL blood*
Product key features
- Highly Efficient CD4+ Cell Isolation: Selectively isolates CD4+ T cells from whole blood with high purity and yield.
- Simple & Fast Protocol: Streamlined process with minimal hands-on time.
- High Purity & Viability: Ensures high-quality isolated CD4+ cells, free from beads and contaminating cells.
- Scalable Protocol: Easily adaptable for different blood volumes to suit specific experimental needs.
- Optimized Reagents: Includes specific conjugates, isolation buffer, and release buffer for optimal isolation and recovery of CD4+ cells.
Product description
The ReadiPrep™ CD4+ Cell Isolation Kit provides a reliable and efficient method for isolating pure, viable CD4+ T cells from whole blood. This kit uses an immunomagnetic separation technique with specific CD4-conjugated beads, ensuring the targeted isolation of CD4+ cells while removing non-CD4+ cells and debris.
The protocol is quick and straightforward, designed to be completed in under 30 minutes with minimal hands-on time. It is optimized for processing up to 30 mL of blood, with all necessary reagents (CD4 conjugate, magnetic spheres, isolation buffer, and release buffer) included in the kit. The isolated CD4+ cells are ideal for downstream applications, including flow cytometry, gene expression analysis, and immunological assays.
The ReadiPrep™ CD4+ Cell Isolation Kit offers high reproducibility and purity, making it an ideal choice for researchers studying immune responses, T cell activation, or conducting clinical trials.
Example protocol
AT A GLANCE
- Take 1mL blood and incubate with 25uL ReadiPrep™ CD4 Conjugate (Component A).
- Incubate for 10 min at RT.
- Add 2mL of 1X isolation buffer and centrifuge at 350g for 15 min.
- Remove the supernatant (remove the same volume of isolation buffer added).
- Add 30uL Magnetic Spheres (Component B). Incubate 10 min at RT.
- Place the tube on a magnetic rack.
- Wait for 2-3 min. Remove supernatant (containing CD4 negative cells)
- Wash beads with 1X Isolation Buffer.
- Add Release Buffer to beads. Incubate for 10 min at RT (with rotating the tube)
- Place the tube in a magnetic rack and collect the supernatant (containing CD4 positive cells).
SAMPLE EXPERIMENTAL PROTOCOL
Prepare 1x Isolation Buffer:
Add 10mL 10X Isolation Buffer (10X) (Component C) to 90mL cell culture water to prepare 1x Isolation buffer.
Blood Sample Preparation:
Take 1 mL whole blood cells with appropriate anticoagulants.
Stain with ReadiPrep™ CD4 Conjugate
Add 25 µL ReadiPrep™ CD4 Conjugate (Component A) to the 1 mL of blood sample.
Incubate for 10 min at RT.
Add 2 mL 1x Isolation Buffer and centrifuge tube at 350g for 15 min at RT.
Remove 2 mL supernatant. Keep 1 mL solution.
Isolate CD4+ Cells:
Add 30 µL Magnetic Spheres (Component B) to above 1 mL cells.
Incubate for 10 min at RT.
Place the tube on a magnetic rack.
Discard supernatant and wash the Magnetic Spheres (with CD4+ cells) with 1 mL of isolation buffer.
Repeat 2-3 times until you get a clear beads solution.
Release CD4+ Cells:
Add 500 µL Elution buffer (Component C).
Incubate 10 min at RT (rotating the tube).
Place the tube on a magnetic rack and collect supernatant containing CD4 positive cells.
To remove any residual beads, place the released CD4+ cells on a magnetic rack for one minute.
Transfer supernatant (CD4+ cells) to a new tube.
Note: This step can be performed twice to get more pure CD4+ positive cells.
The isolated cells are pure, viable, and are free of beads bound to the surface.
CD4+ Cells Storage
Add 500 µL 1x Isolation buffer to the isolated CD4+ cells and centrifuge at 300g for 10 min.
Remove supernatant and suspended cell pellets in any preferred medium.
Store at 4 °C.
Volume of reagents is scalable for specific volume of bloods:
| WBC volume | Component A | Component B | Component C |
| 1 mL | 25 µL | 30 µL | 0.5-1 mL |
References
Authors: Radhi, Noor and Paul, Ayamita and Muelbert, Mariana and Toldi, Gergely
Journal: Journal of human lactation : official journal of International Lactation Consultant Association (2025): 8903344251316491
Authors: Maravelia, Panagiota and Yao, Haidong and Cai, Curtis and Nascimento Silva, Daniela and Fransson, Jennifer and Nilsson, Ola B and Lu, Yong-Chen William and Micke, Patrick and Botling, Johan and Gatto, Francesca and Rovesti, Giulia and Carlsten, Mattias and Sallberg, Matti and Stål, Per and Jorns, Carl and Buggert, Marcus and Pasetto, Anna
Journal: Gut (2025)
Authors: Levin, Noam and Kim, Sanghyun P and Marquardt, Charles A and Vale, Nolan R and Yu, Zhiya and Sindiri, Sivasish and Gartner, Jared J and Parkhurst, Maria and Krishna, Sri and Lowery, Frank J and Zacharakis, Nikolaos and Levy, Lior and Prickett, Todd D and Benzine, Tiffany and Ray, Satyajit and Masi, Robert V and Gasmi, Billel and Li, Yong and Islam, Rafiqul and Bera, Alakesh and Goff, Stephanie L and Robbins, Paul F and Rosenberg, Steven A
Journal: Journal for immunotherapy of cancer (2024)
Authors: Blanco, José-Ramón and Gonzalez-Baeza, Alicia and Martinez-Vicente, Ana and Albendin-Iglesias, Helena and De La Torre, Javier and Jarrin, Inma and González-Cuello, Inmaculada and Cabello-Clotet, Noemí and Barrios-Blandino, Ana-María and Sanjoaquin-Conde, Isabel and Montes-Ramirez, Mª-Luisa and Melus, Estrella and Pérez-Esquerdo, Verónica and Tomas-Jimenez, Cristina and Saumoy-Linares, María and Lopez-Lirola, Ana-Mª and Hidalgo-Tenorio, Carmen and Muelas-Fernandez, Magdalena and Galindo-Puerto, Mª-José and Abadía, Jessica and Manzanares, Eduardo and Segundo-Martin, Cristina and Fernandez-Lopez, Mª-Angeles and Barrios-Vega, María and De Miguel, Marta and Olalla, Julian and ,
Journal: HIV medicine (2024)
Authors: Hagen, Ruth R and Xu, Calvin and Koay, Hui-Fern and Konstantinov, Igor E and Berzins, Stuart P and Kedzierska, Katherine and van de Sandt, Carolien E
Journal: Journal of immunological methods (2024): 113651
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