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The reconstituted Veri-Cells™ Activated (Cytokine) PBMC can be used for immunofluorescent staining and multi-color flow cytometry assays. The cells can be used as controls for cytokine staining with minimal variability across experiments. These cells have been verified to work with commonly tested cell surface (CD3, CD4, CD8, CD19, CD56) as well as cytokine markers such as IL-2, IL-3, IL-4, IL-5, IL-8, IL-13, IL-17, GM-CSF, IFN gamma, and TNF alpha. Some targets such as IL-5 are rare and will require the acquisition of at least 100,000 CD3+ events.
This product contains human cells, a potentially biohazardous material. Blood used in preparation of these samples was tested and found to be negative for antibody and nucleic acid testing against Human Immunodeficiency virus (HIV), Hepatitis B and C virus and HTLV (Human T-Lymphotrophic virus I and II), surface antigen for Hepatitis B (HBsAg) virus and syphilis and West Nile Virus using FDA approved methods. Biological tests are not 100% accurate. Use standard precautions when handling, and treat as if the product is capable of transmitting disease. When handling or disposing, follow precautions described in CDC and FDA recommendations and OSHA Bloodborne Pathogen guidelines.Product Details
- Kit Contents
For Cat. No. 427001:
2 vials of Veri-Cell™ Activated (Cytokine) PBMC (≥1 X 106 cells per vial)
1 vials of 0.7 ml Veri-Cells™ Buffer A
For Cat. No. 427003:
10 vials of Veri-Cell™ Activated (Cytokine) PBMC (≥1 X 106 cells/vial)
5 vials of 0.7 ml Veri-Cells™ Buffer A
Cat. No. 427001: 2 vials each containing ≥ 1 X 106 lyophilized human peripheral blood mononuclear cells (PBMC).
Cat. No. 427003: 10 vials each containing ≥ 1 X 106 lyophilized human peripheral blood mononuclear cells (PBMC).
- The Veri-Cells™ Activated (Cytokine) PBMC kit is prepared from 2 vials of lyophilized human activated PBMCs and a vial of Veri-Cells™ Buffer A. Veri-Cells Activated (Cytokine) PBMC contain lyophilized PBMCs that had been stimulated using Cell Activation Cocktail with Brefeldin A (Cat. No. 423303) for 6 hours.
- Storage & Handling
- Store the kit at 2°C – 8°C upon receipt. Do not open sealed pouch until ready to use. Once reconstituted, the cells may be used for up to five days if properly stored at 2°C – 8°C in the buffer provided.
FC - Quality tested
- Recommended Usage
50 µl per test (after reconstitution with 325 µl of Veri-Cells™ Buffer A)
Note: Veri-Cells™ Buffer A may show precipitation over time, however this is normal and does not affect performance of the buffer.
- Application Notes
Veri-Cells™ Activated (Cytokine) PBMC Kit can be used as a positive control for detecting common surface (CD3, CD4, CD8, CD19, CD56) as well as cytokine markers such as IL-2, IL-3, IL-4, IL-5, IL-8, IL-13, IL-17a, GM-CSF, IFN gamma, and TNF alpha. Assessing the suitability of using this product as a control for end user’s molecule of interest is recommended, as each lot is validated based on the expression levels of markers mentioned above. For best results, Cell Staining Buffer is recommended for any washing steps. Product performance specifications are based on testing with BioLegend reagents and protocols. Deviations from the use of these reagents and procedures have not been assessed and may affect performance.
Protocol for using the Veri-Cells™ Activated (Cytokine) PBMC Kit:
- Prior to reconstitution, warm lyophilized vial(s) of Veri-Cell™ Activated (Cytokine) PBMC at room temperature for 5 minutes
- Add 325µl of Veri-Cell™ Buffer A to the cell pellet. Replace the rubber stopper and mix gently for 5 seconds. Allow at least 5 minutes for the cells to rehydrate prior to staining. Content in each vial is intended for 5 tests
- Optional, fix and permeabilize the cells (Cat. Nos. 420801 and 421002 recommended)
Note: Veri-Cells™ Activated (Cytokine) PBMC do not need to be fixed and permeabilized to achieve cytokine staining. Optional fix/perm steps can be added here to serve as a process control to match experimental sample treatments, if desired.
- Aliquot the required amount of cells into tubes (50 µl=200k cells). Stain with antibodies using our Cell Surface Immunofluorescence Staining Protocol. The cells are ready for acquisition. Cell Staining Buffer (Cat. No. 420201) is recommended for washing Veri-Cells™ Activated (Cytokine) PBMC.
- Wash the cells twice with 2ml of Cell Staining Buffer. The cells are ready for acquisition.
Note: To monitor non-specific staining, we recommend using appropriate isotype controls.
- Biology Area
- Molecular Family
- Gene ID
- Are the cells fixed?
They are treated with our proprietary mixture prior to lyophilization.
- Are there any RBCs in Veri-cells™?
- There may be a few, but the majority of RBCs are not present in Veri-Cells™ preparations.
- Are these cells viable?
- No they are not.
- Are these cells virus/pathogen free?
- These are tested to be free of HIV, HBV, syphilis, and HCV.
- Can a constant percentage of each population be expected in each lot?
- Yes, within each lot, cell populations remain constant.
- Can the cells be activated after rehydration?
- No, they cannot be activated.
- Can they be used to detect intracellular cytokines such as TNF-α or IFNγ?
- Veri-Cells™ PBMC and Veri-Cells™ CD4-Low PBMC will not have detectable TNF-α or IFNγ. Please inquire about a custom made Veri-Cells product at email@example.com.
- How do we QC these cells?
- We QC for T, B, NK markers for Veri-Cells™ PBMC products - CD3, CD19, CD4, CD8, CD56/CD16.
- Is washing required after staining?
- Yes, it is recommended to wash after staining. We recommend using BioLegend's Cell Staining buffer (Cat. No. 420201) for the wash step.