Immunohistochemistry (IHC) was performed on formalin-fixed paraffin-embedded (FFPE) patient sections with an automated staining system (Dako Link48) using TTF1 primary antibodies (ZSGB-Bio Cat: ZM-0270). After deparaffinization and rehydration in xylene and ethanol, antigen retrieval was performed in 1× EDTA retrieval solution (pH 8.0) (E-5134, Sigma) with heating. Inactivation of endogenous peroxidase was performed by adding enough drops of 3% hydrogen peroxide (H324-500, Fisher) to cover the whole section for 10 min and followed by primary antibody incubation at recommended dilution. After 60 min incubation at 37 °C, slides were washed twice with PBS, incubated with goat-anti-rabbit antibody (E046201, Dako) and then developed with DAB substrate (k3468, Dako) for 5–8 min depending on the antibodies. Slides were counterstained with hematoxylin (CTS-1090, Biotechnologies) and were scanned with brightfield pathology microscope at 20× magnification (Aperio CS2, Leica) and processed by ImageScope software (Leica). Information on all other chemicals and reagents used in the article can be found in Supplementary S-2 .
E046201
Manufactured by Agilent Technologies
The E046201 is a high-performance laboratory instrument designed for analytical applications. It provides precise and reliable measurements to support scientific research and product testing.
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2 protocols using e046201
1
Automated Immunohistochemistry for TTF1 in FFPE
2
Automated IHC Staining Workflow
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Immunohistochemistry (IHC) was performed on formalin-xed para n-embedded (FFPE) patient sections with an automated staining system (Dako Link48) using TTF1 primary antibodies (ZSGB-Bio Cat: ZM-0270). After depara nization and rehydration in xylene and ethanol, antigen retrieval was performed in 1x EDTA retrieval solution (pH 8.0) (E-5134, Sigma) with heating. Inactivation of endogenous peroxidase was performed by adding enough drops of 3% hydrogen peroxide (H324-500, Fisher) to cover the whole section for 10 min and followed by primary antibody incubation at recommended working dilution. After 60 mins incubation at 37°C, slides were washed twice with PBS, incubated with goat-anti-rabbit antibody (E046201, Dako) and then developed with DAB substrate (k3468, Dako) for 5-8 mins depending on the antibodies. Slides were counterstained with hematoxylin (CTS-1090, Biotechnologies) and were scanned with bright eld pathology microscope at 20X magni cation (Aperio CS2, Leica) and processed by ImageScope software (Leica). Information on all other chemicals and reagents used in the article can be found in Supplemental S-2.
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