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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 (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).
| Cat # | Size | Price | Quantity Check Availability | Save | ||
|---|---|---|---|---|---|---|
| 426601 | 10 tests | $229 | ||||
| 426603 | 50 tests | $932 | ||||
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.
Kit Contents
- Kit Contents
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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
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ICFC - Quality tested
- Recommended Usage
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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
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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 D57.2.2E); Phospho-p38 (Clone 36/p38); 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
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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
- Phospho-Proteins
- Gene ID
- NA
Related FAQs
- Is washing required after staining?
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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?
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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?
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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?
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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.
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