Veri-Cells™ Phospho PBMC (MAPK/ERK Pathway)

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Regulatory Status
RUO
Other Names
Control Cells
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Product Citations
publications
Veri_Cells_Phospho_PBMC_1_032018
Veri-Cells™ Phospho PBMC (MAPK/ERK pathway) vehicle control (open histogram) or Veri-Cells™ Phospho PBMC (MAPK/ERK pathway) stimulated control (closed histogram) were stained with anti-human phospho S6-Alexa Fluor® 488 (Left), anti-human phospho p38-PE (Center) or anti-human phospho ERK-Alexa Fluor® 647 (Right). The histograms are gated on the lymphocyte population.
  • Veri_Cells_Phospho_PBMC_1_032018
    Veri-Cells™ Phospho PBMC (MAPK/ERK pathway) vehicle control (open histogram) or Veri-Cells™ Phospho PBMC (MAPK/ERK pathway) stimulated control (closed histogram) were stained with anti-human phospho S6-Alexa Fluor® 488 (Left), anti-human phospho p38-PE (Center) or anti-human phospho ERK-Alexa Fluor® 647 (Right). The histograms are gated on the lymphocyte population.
  • Veri_Cells_Phospho_PBMC_2_032018
    Veri-Cells™ Phospho PBMC (MAPK/ERK pathway) scatter pattern. Veri-Cells™ Phospho PBMC (MAPK/ERK pathway) Vehicle Control (Left) and Veri-Cells™ Phospho PBMC (MAPK/ERK pathway) Stimulated Control (Right).
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426601 10 tests 229 CHF
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426603 50 tests 932 CHF
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Description

The reconstituted human PBMCs can be used for intracellular flow cytometry assays. They can also be used as a process control to ensure effective permeabilization when performing intracellular staining using True-Phos™ Perm Buffer.

This product contains human cells, a potentially biohazardous material. Blood used in preparation of these samples was tested and found to be negative, using FDA approved methods, 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. 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
Technical Data Sheet (pdf)

Kit Contents

Kit Contents

Cat. No. 426601

  • 2 vials of Veri-Cells™ Phospho PBMC (MAPK/ERK pathway) Vehicle Treated Cells
  • 2 vials of Veri-Cells™ Phospho PBMC (MAPK/ERK pathway) Stimulated Cells
  • 1 vial of Veri-Cells™ Reconstitution Buffer A Plus


Cat. No. 426603

  • 10 vials of Veri-Cells™ Phospho (MAPK/ERK pathway) Vehicle Treated Cells
  • 10 vials of Veri-Cells™ Phospho (MAPK/ERK pathway) Stimulated Cells
  • 5 vials of Veri-Cells™ Reconstitution 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
Each kit contains an equal number of vials of unstimulated (vehicle treated) and stimulated Veri-Cells™ Phospho PBMC (MAPK/ERK pathway) cells (2 vials each for 10 tests, and 10 vials each for 50 tests), as well as Veri-Cells™ Buffer A Plus (1 vial for 10 tests and 5 vials for 50 tests). Each vial of Veri-Cells™ Phospho PBMC (MAPK/ERK pathway) contains approximately 1 x 106 lyophilized human peripheral blood mononuclear cells (PBMC).
Preparation
The Veri-Cells™ Phospho PBMC (MAPK/ERK pathway) Kit is prepared by stimulating human PBMCs with the Cell Activation Cocktail (Cat. No. 423301) for 15 minutes. The lyophilized cells can be reconstituted with Veri-Cells™ Buffer A Plus and permeablized with True-Phos™ Perm Buffer (Cat. No. 425401).
Storage & Handling
Store the kit at 2°C - 8°C upon receipt. Do not open the sealed pouch until ready to use. The cells may be used for up to 5 days if properly stored at 2°C - 8°C in the buffer provided and processed according to the defined protocol.
Application

ICFC - 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

The Veri-Cells Phospho PBMC (MAPK/ERK pathway) Kit can be used as both positive and negative staining controls for members of the RAS mitogen-activated kinase (MAPK/ERK) pathway such as phospho-S6, phospho-p38 and phospho-ERK 1/2. Additional intracellular targets may be detectable outside of those detailed on the CoA. Once reconstituted, the Veri-Cells™ Phospho PBMC (MAPK/ERK pathway) are processed and stained per the protocol detailed below. Deviations from the use of these reagents and procedures have not been assessed and may affect performance.

Cell subset

Acceptance Criteria, % positive of lymphocyte gate

Phospho-S6

60-100

Phospho-p38

30-100

Phospho-ERK

70-100


Note on indicated ranges: Percent positive values are obtained based on gates set on unstained cells. The antibodies used for testing were the following: Phospho-S6 (Clone A17020B); Phospho-p38 (Clone A16016A); Phospho-ERK (Clone 4B11B69). 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 Phospho PBMC (MAPK/ERK Pathway). Percent positive is based on gates set on unstained cells. The Certificate of Analysis specific values are provided only as a reference. Variability in antibodies used, instruments, staining conditions and other factors may influence end-user values. It is recommended that individual laboratories establish its own control values and ranges for each parameter based on laboratory specific conditions and protocols.

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.

Product Citations
  1. de Andrade Pereira B, et al. 2013. Gene Ther. 556:20. PubMed

Antigen Details

Biology Area
Immunology
Molecular Family
Phospho-Proteins
Gene ID
NA

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.

Go To Top Version: 6    Revision Date: 05.02.2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

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