Liver tissue was fixed in ice-cold 10% formalin and paraffin embedded tissue sections (5μm) were used for IHC. Antigen retrieval was performed in 1×citrate buffer in microwave pressure cooker. We used 1/200 dilution of BrdU plus Phospho-histone H3 antibodies (phospho-histone H3:Cell signalling #9701; BrdU BD). Phospho-histone H3 (P-histone H3) was detected using a biotinylated goat Anti-Rabbit antibody (Dako) and the AB reagents from Vectastain (Vector labs). The fluorescent signal was provided using the Fluorescein Tyramide Signal Amplification (TSA) Systems (Perkin elmer). BrdU antibody was detected using a goat anti mouse 1/200 Alexa 568 antibody (Molecular Probes life technologies). Cells were counterstained with Dapi (500ng/ml) and slides were mounted with mowiol. Pictures were taken using a Leica DMI6000 time lapse microscope.
Fluorescein tyramide signal amplification systems
Manufactured by PerkinElmer
Fluorescein Tyramide Signal Amplification (TSA) Systems are a set of reagents and protocols designed to enhance and amplify fluorescent signals in immunohistochemistry and in situ hybridization applications. The core function of the TSA systems is to provide a signal amplification method that increases the sensitivity of target detection.
Automatically generated - may contain errors
2 protocols using fluorescein tyramide signal amplification systems
1
Multiparametric Analysis of Proliferation
2
Multiparametric Analysis of Proliferation
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Liver tissue was fixed in ice-cold 10% formalin and paraffin embedded tissue sections (5μm) were used for IHC. Antigen retrieval was performed in 1×citrate buffer in microwave pressure cooker. We used 1/200 dilution of BrdU plus Phospho-histone H3 antibodies (phospho-histone H3:Cell signalling #9701; BrdU BD). Phospho-histone H3 (P-histone H3) was detected using a biotinylated goat Anti-Rabbit antibody (Dako) and the AB reagents from Vectastain (Vector labs). The fluorescent signal was provided using the Fluorescein Tyramide Signal Amplification (TSA) Systems (Perkin elmer). BrdU antibody was detected using a goat anti mouse 1/200 Alexa 568 antibody (Molecular Probes life technologies). Cells were counterstained with Dapi (500ng/ml) and slides were mounted with mowiol. Pictures were taken using a Leica DMI6000 time lapse microscope.
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