Veri-Cells™ Activated (Cytokine) PBMC

Kit Contents

Kit Contents

Cat. No. 427001

• 2 vials of Veri-Cell™ Activated (Cytokine) PBMC (≥1 X 10⁶ cells per vial) 
• 1 vial of 0.7 mL Veri-Cells™ Buffer A Plus

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 Plus
   

Veri-Cells™ Buffer A Plus is an improved version of the original Veri-Cells™ Buffer A to improve staining performance with tandem dyes.

Product Details

Formulation

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).

Preparation

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 Plus. 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.

Application

FC - Quality tested

Recommended Usage

50 µL per test (after reconstitution with 325 µl of Veri-Cells™ Buffer A Plus)

Once reconstituted, let the cells sit for 15 minutes at room temperature prior to staining.

Veri-Cells™ Buffer A Plus 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 verified 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.

Antibodies Used for Product Validation
Target	Clone	Fluorophore
CD3	SK7	FITC
IL-13	JES10-5A2	PE/Cyanine7
GM-CSF	BVD2-21C11	Pacific Blue™
TNF-alpha	Mab11	PerCP/Cyanine5.5
IL-2	MQ1-17H12	APC/Cyanine7
IL-8	E8N1	APC
IL-17a	BL168	Brilliant Violet 421™
IFN-gamma	4S.B3	APC
IL-5	TRFK5	APC
IL-4	MP4-25D2	PE/Cyanine7
IL-3	BVD3-1F9	PE

 

Cell subset	Acceptance Criteria, % positive of lymphocyte gate
CD3+IL-2+	10-40
CD3+IL-3+	0.2-5
CD3+ IL-4+	0.1-3
CD3+ IL-5+	0.2-3
CD3+IL-8+	0.1-5
CD3+IL-13+	0.2-5
CD3+IL-17A+	0.1-2
CD3+IFN gamma+	5-30
CD3+TNF alpha+	10-60
CD3+GM-CSF+	1-15

Note on indicated ranges: The antibodies used for testing were the following: CD3 APC/Cyanine7 (clone SK7), IL-2 APC/Cyanine7 (clone MQ1-17H12), IL-3 PE (clone BVD3-1F9), IL-4 PE/Cyanine7 (clone MP4-25D2), IL-5 APC (clone TRFK5), IL-8 PE/Cyanine7 (clone E8N1), IL-13 APC (clone JES10-5A2), IL-17a Brilliant Violet™ 421 (clone BL168), GM-CSF Pacific Blue™ (clone BVD2-21C11), TNF-α PerCP/Cyanine5.5 (clone MAB11), and IFN-ʎ APC (clone 4S.B3). Variability in antibodies used, instruments, staining conditions and other factors may influence end-user values. The Certificate of Analysis specific values are provided as a reference. It is recommended that individual laboratories establish its own control values and ranges for each parameter based on laboratory specific conditions and protocols.

Please use our CoA Look-Up Tool to find the expected lot-specific frequencies of certain cell populations included within each lot of Veri-Cells™ Activated (Cytokine) PBMC. 

Additional Product Notes

Veri-Cells™ Buffer A Plus is an improved version of the original Veri-Cells™ Buffer A to improve staining performance with tandem dyes.

Antigen Details

Biology Area

Immunology

Molecular Family

Cytokines/Chemokines

Gene ID

NA

Documentation

• Technical data sheet
• Certificate of Analysis
• Material Safety Data Sheet

Reviews

Related Protocols

• Protocol for using Veri-Cells™

Related Pages & Pathways

Related FAQs

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.

Can the cells be activated after rehydration?

No, they cannot be activated.

Are these cells virus/pathogen free?

These are tested to be free of HIV, HBV, syphilis, and HCV.

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.

Can a constant percentage of each population be expected in each lot?

Yes, within each lot, cell populations remain constant.

Are the cells fixed?

They are treated with our proprietary mixture prior to lyophilization.

Can they be used to detect intracellular cytokines such as TNF-α or IFN-γ?

For this, use Veri-Cells™ Activated (Cytokine) PBMC which have been activated with Cell Activation Cocktail (with Brefeldin A) containing PMA and Ionomycin. These have been validated to stain positive for activation-induced cytokines including TNF-α and IFN-γ. Veri-Cells™ PBMC and Veri-Cells™ CD4-Low PBMC will not have detectable TNF-α or IFN-γ.

How do we QC these cells?

Each Veri-Cells™ product is tested with a panel of antibodies to ensure expression of these markers falls within the expected range.  For more information regarding the exact markers used for each product, please contact technical service.

Certificate of Analysis