AKOYA PhenoCode Discovery Tissue Architecture Human Protein Module User Guide
- August 9, 2024
- AKOYA
Table of Contents
AKOYA PhenoCode Discovery Tissue Architecture Human Protein Module
Specifications
-
STORAGE
- Antibodies : 4°C
- Reporters : -20°C
See PhenoCycler-Fusion User Guide (Doc# PD-000011) for details.
-
STABILITY
See the 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).
Product Information
The PhenoCode Discovery Tissue Architecture Human Protein Module is
designed for tissue staining and analysis. It contains a selection of
conjugated antibodies and reporters for specific targets.
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
Optimized for the PhenoCycler-Fusion system. Refer to the PhenoCycler-Fusion User Guide (Doc# PD-000011) for the staining protocol. -
Dilution
Dilute all antibodies 1:200, except for Vimentin and Beta-actin which should be diluted 1:400. -
Cycle Configuration on PhenoCycler-Fusion
The recommended run cycle configuration on the PhenoCycler-Fusion system for imaging tonsil and lung cancer tissue is provided in the manual. Exposure times for each target are listed accordingly.
The human FFPE lung cancer tissue section was stained with the PhenoCode Discovery Tissue Architecture 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, except for Vimentin and Beta-actin, which were diluted 1:400.
The PhenoCode™ Discovery Tissue Architecture Human Protein Module enables the detection of 10 markers on multiple tissues using the PhenoCycler®-Fusion system. It is intended to help researchers understand the distribution of vasculature and look for epithelial-to-mesenchymal transition in the tumor microenvironment. It has been tested on cancer tissue.
| Target | Biological Relevance |
|---|---|
| E-cadherin | Epithelial cells |
| SMA | Vascular structures, fibroblasts, smooth muscle |
| Vimentin | Cytostructures |
| Collagen IV | Extracellular matrix (ECM) |
| CD34 | Vascular structures |
| CD31 | Vascular structures |
| Beta-catenin1 | Cell adhesion, Wnt signaling |
| Beta-actin | Cytoskeleton, housekeeping gene |
| Podoplanin | Lymphatics |
| Caveolin | Integral membrane protein |
Contents of PhenoCode Discovery Tissue Architecture Human Protein Module
The PhenoCode Discovery Tissue Architecture Human Protein Module contains the
following conjugated antibodies and reporters:
Target
|
Catalog #
|
Clone ID
|
Barcode
|
Reporter
| Dilution Tonsil| Dilution Cancer
---|---|---|---|---|---|---
E-cadherin|
4250021| AKYP0057|
BX014| Atto 550-RX014| 1:200| 1:200
SMA| 4450049|
AKYP0081| BX013| Alexa Fluor™ 750-RX013| 1:200| 1:200
Vimentin| 4450050|
AKYP0082| BX022| Alexa Fluor™ 750-RX022| 1:400| 1:400
Collagen IV|
4550122| AKYP0083|
BX042| Alexa Fluor™ 647-RX042| 1:200| 1:200
CD34| 4550133|
AKYP0088| BX025| Alexa Fluor™ 647-RX025| 1:200| 1:200
CD31| 4250104|
AKYP0047| BX001| Atto 550-RX001| 1:200| 1:200
Beta-catenin1|
4250091| AKYP0068|
BX096| Atto 550-RX096| 1:200| 1:200
Beta-actin|
4450092| AKYP0072|
BX117| Alexa Fluor™ 750-RX117| 1:500| 1:400
Podoplanin|
4250094| AKYP0007|
BX121| Atto 550-RX121| 1:200| 1:200
Caveolin| 4450084|
AKYP0115| BX086| Alexa Fluor™ 750-RX086| 1:200| 1:200
Cycle Configuration on PhenoCycler-Fusion
The PhenoCode Discovery Tissue Architecture Human Protein Module was validated
using the following run 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. Recommended
exposure times are listed for imagining tonsil and lung cancer tissue.
Exposure times are for PhenoCycler- Fusion only.
TONSIL
TISSUE
Cycle Order
|
Atto 550
|
Alexa Fluor 647
|
Alexa Fluor 7 50
---|---|---|---
Target-Barcode| Exposure Time
(ms)
| Target-Barcode|
Exposure Time (ms)
| Target-Barcode|
Exposure Time (ms)
1| E-cadherin-BX014| 100| Collagen IV-BX042| 125| Vimentin-BX022| 60
2| CD31-BX001| 50| CD34-BX025| 125| SMA-BX013| 80
3| Beta-catenin1-BX096| 100| —| —| Beta-actin-BX117| 50
4| Podoplanin-BX121| 100| —| —| Caveolin-BX086| 50
CANCER TISSUE
Cycle Order
| Atto 550| Alexa Fluor 647| Alexa Fluor 7 50
---|---|---|---
Target-Barcode| Exposure Time
(ms)
| Target-Barcode|
Exposure Time (ms)
| Target-Barcode|
Exposure Time (ms)
1| E-cadherin-BX014| 100| Collagen IV-BX042| 125| Vimentin-BX022| 60
2| CD31-BX001| 50| CD34-BX025| 125| SMA-BX013| 80
3| Beta-catenin1-BX096| 100| —| —| Beta-actin-BX117| 50
4| Podoplanin-BX121| 100| —| —| Caveolin-BX086| 50
HUMAN FFPE TONSIL SECTION
Human FFPE tonsil section was stained with the PhenoCode Discovery Tissue Architecture Human Protein Module and imaged on the PhenoCycler-Fusion system. Representative imaging regions show Vimentin (cyan), Caveolin (magenta), Podoplanin (yellow), E-cadherin (red), CD31 (green),Collagen IV (white) and DAPI (blue). Antigen retrieval was performed using AR9 (Akoya, Part# AR900250ML). All antibodies shown here were diluted 1:200 except for Vimentin, which was diluted 1:400.
**HUMAN FFPE LUNG CANCER SECTION
**
Human FFPE lung cancer section was stained with the PhenoCode Discovery Tissue Architecture Human Protein Module and imaged on the PhenoCycler-Fusion system. Representative imaging regions show Caveolin (magenta), E-cadherin (red), Podoplanin (green), Vimentin (orange), SMA (cyan), CD31 (yellow), and Collagen IV (dark purple). Antigen retrieval was performed using AR9 (Akoya, Part# AR900250ML). All antibodies shown here were diluted 1:200 except for Vimentin, which was diluted 1:400.
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For Research Use Only. Not for use in diagnostic procedures.
© 2024 Akoya Biosciences, Inc. All rights reserved. All trademarks are the property of Akoya Biosciences unless otherwise specified. PD-000067 Rev. B
FAQ
Q: What species does the PhenoCode Discovery Tissue Architecture Human
Protein Module react with?
A: The module is designed for use with human tissue samples.
Q: How should I perform antigen retrieval for staining with this module?
A: Use AR9 (Akoya, Part# AR900250ML) for antigen retrieval according to the
instructions provided in the manual.
Q: What is the recommended dilution for the antibodies in this module?
A: Dilute all antibodies at 1:200, except for Vimentin and Beta-actin which
should be diluted at 1:400.