Formalin-fixed paraffin-embedded (FFPE) tumor samples from patients with HNSCC were stained using Opal 7-Color Automation IHC Kit-50 Slide according to manufacturer’s instructions (Akoya Biosciences). Briefly, antigen retrieval was performed using AR6 buffer in a microwave at 100% power for 1 min and 10% power for 15 min. Slide staining was performed in three cycles: (1) first cycle, antihuman CD20 primary antibody (1:400) and Opal 540 (1:50); (2) second cycle, antihuman CD8 primary antibody (1:100) and Opal 650 (1:50); (3) third cycle, antihuman pan-cytokeratin primary (1:500) and Opal 690 (1:50). Slides were stained with DAPI for 5 min, washed and mounted and scanned using the Vectra 3 system and Phenochart software. Acquired MSI images were unmixed and analyzed by inForm 2.0 software. Detailed antibody information is included in online supplemental table 8 .
Opal 7 color automation ihc kit 50 slide
Manufactured by Akoya Biosciences
The Opal 7-Color Automation IHC Kit-50 Slide is a laboratory equipment product from Akoya Biosciences. It is designed for performing immunohistochemistry (IHC) experiments with up to 7 different markers on a single slide in an automated manner.
Automatically generated - may contain errors
Lab products found in correlation
Bond rx, by Leica (1 mentions)
Spectral dapi, by Akoya Biosciences (1 mentions)
Prolong diamond mounting media, by Thermo Fisher Scientific (1 mentions)
Cd68 clone kp1, by Agilent Technologies (1 mentions)
Pd l1 clone e1l3n, by Cell Signaling Technology (1 mentions)
Bond wash, by Leica (1 mentions)
Antibody ab diluent, by Akoya Biosciences (1 mentions)
Hrp reactive opal fluorescent reagents, by Akoya Biosciences (1 mentions)
2 protocols using opal 7 color automation ihc kit 50 slide
1
Multiplexed Immunohistochemistry of HNSCC Tumors
2
Multiplex Immunofluorescence for HNSCC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Twenty-three formalin-fixed paraffin-embedded human HNSCC samples were stained using the Opal 7-Color Automation IHC Kit-50 Slide according to manufacturer’s instructions (Akoya Biosciences) using the following antibodies and antigen retrieval processes: with antibodies against PD-L1 (clone E1L3N, Cell Signaling Technologies cat. 13684S, Opal 620), CD11c (clone 5D11, Sigma Cell Marque cat. 111M-15, Opal 650), CD68 (clone KP1, Dako cat. M0814, Opal 690), and pan-CK (clone AE1/AE3, Dako cat. M351501-2, Opal 540) on a Bond RX autostainer (Leica Biosystems). Slides were dewaxed (Leica), heat treated in ER2 or ER1 antigen retrieval buffer depending on the antibody for 20 min at 93 °C (Leica), blocked in Antibody (Ab) Diluent (Akoya Biosciences), incubated for 30 min with the primary Ab, 10 min with horseradish peroxidase (HRP)-conjugated secondary polymer (anti-rabbit and anti-mouse, Akoya Biosciences), and 10 min with HRP-reactive OPAL fluorescent reagents (Akoya Biosciences). Slides were washed between staining steps with Bond Wash (Leica) and stripped between each round of staining with heat treatment in antigen retrieval buffer. After the final heat treatment in antigen retrieval buffer, the slides were stained with spectral DAPI (Akoya Biosciences), and coverslipped with Prolong Diamond mounting media (Thermo Fisher).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!