AKOYA AKYP0098 Pheno Code Discovery Lymphocyte Profiling Human Protein Modul User Guide
- September 14, 2024
- AKOYA
Table of Contents
AKOYA AKYP0098 Pheno Code Discovery Lymphocyte Profiling Human Protein
Modul
Specifications:
- Product Name: PhenoCode Discovery Lymphocyte Profiling Human
- Protein Module
- Species Reactivity: Human
- Tissue Type: FFPE
- System Compatibility: PhenoCycler-Fusion system
Product Usage Instructions
Stability :
Check the expiration date of each antibody and reporter tube before use.
Antigen Retrieval:
Use AR9 (Akoya, Part# AR900250ML) for antigen retrieval.
System Compatibility:
The panel module is optimized for the PhenoCycler-Fusion system. Refer to
the PhenoCycler-Fusion User Guide (Doc# PD-000011) for the tissue staining
protocol.
Biological Relevance:
The PhenoCode Discovery Lymphocyte Profiling Human Protein Module targets
various cell types including T cells, B cells, and NK cells. It provides
insights into T cell exhaustion, regulatory T cells, activated T cells/NK
cells, B cells, Wnt signaling, and more.
FAQ :
-
Q: What species is the PhenoCode Discovery Lymphocyte Profiling Human Protein Module optimized for?
A: The module is optimized for human species reactivity. -
Q: How should I handle antigen retrieval for this product?
A: Use AR9 (Akoya, Part# AR900250ML) for antigen retrieval as specified in the instructions. -
Q: What is the recommended dilution for the antibodies in the module?
A: The antibodies should be diluted at a ratio of 1:200 before use.
TECHNICAL DATA GUIDELINES | Panel for PhenoCode Discovery
PhenoCode Discovery Lymphocyte Profiling Human Protein Module
PRODUCT INFORMATION
STORAGE
- Antibodies: 4°C
- Reporters: -20°C*
- See PhenoCycler-Fusion User Guide (Doc# PD-000011) for details.
STABILITY
See expiration date of each antibody and reporter tube.
ANTIGEN RETRIEVAL
AR9 (Akoya, Part# AR900250ML)
SPECIES REACTIVITY
Human
TISSUE TYPE
FFPE
SYSTEM COMPATIBILITY
The panel module has been optimized for the PhenoCycler-Fusion system.
Protocol for tissue staining can be found in the PhenoCycler-Fusion User Guide
(Doc# PD-000011).
Human FFPE lung cancer tissue section was stained with the PhenoCode Discovery Lymphocyte Profiling Human Protein Module and imaged on the PhenoCycler-Fusion system. Antigen retrieval was performed using AR9 (Akoya, Part# AR900250ML). All antibodies were diluted 1:200.
The PhenoCodeTM Discovery Lymphocyte Profiling Human Protein Module enables detection of 8 markers on multiple tissues using the PhenoCycler®-Fusion system. It is intended to help researchers understand the types and states of T cells and B cells in the tumor microenvironment.
| Target | Biological Relevance |
|---|---|
| TOX | T cell exhaustion |
| FOXP3 | Regulatory T cells |
| Granzyme B | Activated T cells/NK cells |
| CD21 | B cells |
| CD79a | B cells |
| TCF-1 | Wnt signaling |
| CD107a | CD8+ and NK cells; lysosomal marker |
| CD38 | Plasma B cells |
Contents of PhenoCode Discovery Immune Profiling Human Protein Core
The PhenoCode Discovery Tissue Architecture Human Protein Module contains the following conjugated antibodies and reporters. Recommended starting dilutions are listed for staining tonsil and lung cancer tissue on the PhenoCycler- Fusion system. Further optimization may be needed depending on the tissue.
Target| Catalog #| Clone ID| Barcode| Reporter|
Dilution Tonsil| Dilution Cancer
---|---|---|---|---|---|---
TOX| 4250067|
AKYP0098| BX060| Atto 550-RX060| 1:200| 1:200
FOXP3| 4550071|
AKYP0102| BX031| Alexa FluorTM 647 -RX031| 1:200| 1:200
Granzyme B|
4550131| AKYP0086|
BX041| Alexa FluorTM 647-RX041| 1:200| 1:200
CD21| 4450100|
AKYP0061| BX032| Alexa FluorTM 750-RX032| 1:200| 1:200
CD79a| 4250103|
AKYP0109| BX090| Atto 550-RX090| 1:200| 1:200
TCF-1| 4550068|
AKYP0099| BX061| Alexa FluorTM 647-RX061| 1:200| 1:200
CD107a| 4450101|
AKYP0004| BX006| Alexa FluorTM 750-RX006| 1:200| 1:200
CD38| 4450099|
AKYP0110| BX035| Alexa FluorTM 750-RX035| 1:200| 1:200
Cycle Configuration on PhenoCycler-Fusion
The PhenoCode Discovery Immune Profiling Human Protein Core was run using the following cycle configuration on the PhenoCycler-Fusion system using standard recommendations for nuclear stain and blank cycles. The order and cycle configuration of markers can be changed as needed.
Cycle Order| Atto 550| Alexa Fluor 647| Alexa Fluor
750
---|---|---|---
1| TOX| FOXP3| CD21
2| CD79a| Granzyme B| CD38
3| | TCF-1| CD107a
Dilution and Exposure Time Recommendations
The following table indicates recommended starting dilutions and exposure times for staining and imaging tonsil and cancer tissue on the PhenoCycler- Fusion system. Further optimization may be needed depending on the tissue. Exposure times are for PhenoCycler-Fusion only.
Target
| Tonsil| Cancer
---|---|---
Dilution| Exposure Time (ms)| Dilution| Exposure Time (ms)
TOX| 1:200| 125| 1:200| 125
FOXP3| 1:200| 125| 1:200| 125
Granzyme B| 1:200| 125| 1:200| 125
CD21| 1:200| 125| 1:200| 125
CD79a| 1:200| 125| 1:200| 125
TCF-1| 1:200| 125| 1:200| 125
CD107a| 1:200| 125| 1:200| 125
CD38| 1:200| 125| 1:200| 125
HUMAN FFPE TONSIL SECTION
Human FFPE tonsil section was stained with the PhenoCode Discovery Immune Profiling Human Protein Core and imaged on the PhenoCycler-Fusion system. Representative imaging regions showing FOXP3 (cyan), CD21 (green), TOX (yellow), Granzyme B (red), CD38 (orange), and DAPI (Blue). Antigen retrieval was performed using AR9 (Akoya, Part# AR900250ML). All antibodies were diluted 1:200.
DAPI CD21 FOXP3 TOX CD38
Granzyme B
HUMAN FFPE LUNG CANCER SECTION
Human FFPE lung cancer section was stained with the PhenoCode Discovery Immune
Profiling Human Protein Core and imaged on the PhenoCycler-Fusion system.
Representative imaging regions showing TCF-1(magenta), Granzyme B (red),
CD107a (green), CD38 (orange), FOXP3 (cyan), CD79a (yellow) and DAPI (blue).
Antigen retrieval was performed
using AR9 (Akoya, Part#
DAPI CD107a FOXP3 CD79a CD38
Granzyme B TCF-1
| AR900250ML). All antibodies were diluted 1:200.
Human FFPE lung cancer section was stained with the PhenoCode Discovery Immune Profiling Human Protein Core and imaged on the PhenoCycler-Fusion system. Representative imaging regions showing TCF-1(magenta), Granzyme B (red), CD107a (green), CD38 (orange), FOXP3 (cyan), CD79a (yellow) and DAPI (blue). Antigen retrieval was performed using AR9 (Akoya, Part# AR900250ML). All antibodies were diluted 1:200.
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For Research Use Only. Not for use in diagnostic procedures.
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PD-000056 Rev A