With pilot study, we selected biomarkers that differed more than 4 fold in mRNA levels in TRG4 and TRG1. Epidermal growth factor receptor (EGFR), C-ern and Ki67 and Krüppel-like factor 5 (KLF5), known as a transcriptional factor acting down-stream of RAS-MAPK cascade, were selected [15 (link)16 (link)17 (link)18 (link)]. Biopsy slides were blocked with 5% bovine serum albumin in phosphate buffered saline (PBS) and primary antibodies (Abcam, Ltd., Cambridge, United Kingdom) were applied at a 1:50 dilution, followed by incubation with biotinylated anti-rabbit secondary antibody (Dianova, Hamburg, Germany, at a dilution of 1:50). With EGFR, C-ern, the intensity immunostaining was scored as: 1+ (weak), 2+ (moderate), and 3+ (intense). With Ki67 and KLF5, the percentage of positive tumor cells and staining intensity were then multiplied to produce a weighted score for each case ranging from 0 to 12; “low expression” was classified as a score of 5 or below and “high expression” as a score of 6 or above.
Biotinylated anti rabbit secondary antibody
Manufactured by Dianova
Sourced in Germany
Biotinylated anti-rabbit secondary antibody is a laboratory reagent used to detect the presence of rabbit primary antibodies in various immunoassays. It serves as a detection tool, binding to the rabbit primary antibodies and enabling their visualization or further processing.
Automatically generated - may contain errors
Lab products found in correlation
Tsa cy3 system, by PerkinElmer (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Dfc360 fx, by Leica (1 mentions)
Dmi4000b microscope, by Leica (1 mentions)
Anti β catenin, by Cell Signaling Technology (1 mentions)
Anti cleaved caspase 8, by Cell Signaling Technology (1 mentions)
Anti villin, by Santa Cruz Biotechnology (1 mentions)
Anti caspase 8, by Cell Signaling Technology (1 mentions)
2 protocols using biotinylated anti rabbit secondary antibody
1
Biomarker Analysis in Tumor Samples
2
Comprehensive Histopathological and Immunofluorescence Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Histopathological examinations were performed after Mayer’s H&E staining. Immunofluorescence staining was performed on formalin-fixed paraffin-embedded tissue by using the Tyramide Signal Amplification (TSA) Cy3 system (Perkin&Elmer) according to the manufacturer’s protocol. The following primary antibodies were used: Anti-Villin (Santa Cruz), anti-β-Catenin (Cell Signaling), anti-Caspase-8 (Cell Signaling) and anti-Cleaved Caspase-8 (Cell Signaling). A biotinylated anti-rabbit secondary antibody from Dianova was used. Nuclei were counterstained with Hoechst 33342 (Invitrogen). For cell death analysis the In Situ Cell Death Detection Kit for TUNEL (TdT-mediated dUTP nick end labelling) from Roche was used. Bright-field and fluorescence pictures were taken by using the DMI4000 B microscope (Leica) in combination with a LEICA DFC360 FX or LEICA DFC420C camera.
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