Tissue was fixed with 4% formaldehyde in PBS for 48h and sent to VAI histology core for embedding. Deparaffinization and antigen retrieval were performed on the Dako PT link platform using Dako High pH retriever buffer for 20 minutes at 97°C. Staining was performed utilizing Dako Autostainer Link 48, and Dako Rabbit Polymer HRP as secondary antibody for 20 minutes following primary antibody incubation for 30 minutes. DAB detection was performed using Dako EnVision Flex Chromagen for 10 minutes and Dako Flex Hematoxylin for 5 minutes. Aperio scanning of slides was performed utilizing Leica Aperio AT2 system.
High ph retriever buffer
Manufactured by Agilent Technologies
High pH retriever buffer is a laboratory solution used to prepare samples for immunohistochemical (IHC) analysis. It is designed to facilitate the retrieval of target antigens that may have been masked or altered during the fixation process, enabling more accurate detection and visualization during IHC staining procedures.
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Lab products found in correlation
3 protocols using high ph retriever buffer
1
Tissue Immunohistochemistry with DAB Detection
2
Immunohistochemical Staining Protocol
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Tissue was fixed with 4% formaldehyde in PBS for 48h and sent to VAI histology core for embedding. Deparaffinization and antigen retrieval performed on Dako PT link platform using Dako High pH retriever buffer for 20 min at 97°C. Staining performed utilizing Dako Autostainer Link 48, utilizing Dako Rabbit Polymer HRP for secondary for 20min following primary antibody incubation for 30min. DAB detection performed using Dako EnVision Flex Chromagen for 10min and Dako Flex Hematoxylin for 5min. Aperio scanning of slides was performed utilizing Leica Aperio AT2 system.
3
Immunohistochemical Quantification of BAT
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Tissue was fixed with 4% formaldehyde in PBS for 48h and sent to VAI histology core for embedding. Deparaffinization and antigen retrieval performed on Dako PT link platform using Dako High pH retriever buffer for 20 minutes at 97 degrees C. Staining performed utilizing Dako Autostainer Link 48, utilizing Dako Rabbit Polymer HRP for secondary for 20 minutes following primary antibody incubation for 30 minutes. DAB detection performed using Dako EnVision Flex Chromagen for 10 minutes and Dako Flex Hematoxylin for 5 minutes. Aperio scanning of slides was performed utilizing Leica Aperio AT2 system.
ImageJ software (NIH, Bethesda, MD) was used to quantitate the number of nuclei that were stained in each experimental condition. Positively stained nuclei were divided by the total number of nuclei to yield the percent of positive nuclei in BAT.
ImageJ software (NIH, Bethesda, MD) was used to quantitate the number of nuclei that were stained in each experimental condition. Positively stained nuclei were divided by the total number of nuclei to yield the percent of positive nuclei in BAT.
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