rabbit anti human her2 monoclonal antibody  (Cell Signaling Technology Inc)

Journal: medRxiv

Article Title: Validation and Prospective Testing of a High Sensitivity, Quantitative Analytic Assay for HER2 on Histopathology Slides

doi: 10.1101/2025.05.06.25327097

Figure Lengend Snippet: A histogram of 316 measurements of HER2 from the breast cancer core biopsy cases in the prospective trial sorted from lowest to highest and color coded by IHC scores (2+/N means a score of 2+ with reflex FISH testing showing no ERBB2 gene amplification). The LOD and LOQ values for this assay are inset in the lower left.

Article Snippet: By loading one control CMA slide together with every 9 tissue slides in each BOND slide tray, HS-HER2 QIF staining was carried out by the following protocol; deparaffinization with BOND dewax solution (AR9222), antigen retrieval with BOND HIER epitope retrieval solution 2 (AR9640) at 97°C for 20 minutes, blocking with ReadyProbes Endogenous HRP & AP blocking solution (R37629, Invitrogen) for 10 minutes and with BSA for 30 minutes, 1 hour-incubation with primary rabbit monoclonal HER2 antibody (clone 29D8, 2165, IgG, Cell Signaling) at optimal concentration of 1ug/ml mixed together with 1:100 concentration of pan-CK (Clones AE1/AE3,M3515, Dako), amplification with Rabbit Envision+ System – HRP labelled polymer anti-Rabbit (K400311-2, Dako) mixed together with 1:100 dilution of a green-fluorescent Alexa Fluor 546 Goat-anti-Mouse IgG (H+L)Cross-Adsorbed Secondary Antibody (A11003, Invitrogen) for 1 hour, staining with 1:50 dilution of a red-fluorescent Tyramide Signal Amplification (TSA) Cyanin 5 (SAT705A001EA, Akoya Biosciences) for 10 minutes and nuclear staining with 1:500 dilution of a blue-fluorescent 4′,6-diamidino-2-phenylindole (DAPI) for 10 minutes.

Techniques: Amplification

Journal: medRxiv

Article Title: Validation and Prospective Testing of a High Sensitivity, Quantitative Analytic Assay for HER2 on Histopathology Slides

doi: 10.1101/2025.05.06.25327097

Figure Lengend Snippet:

Article Snippet: By loading one control CMA slide together with every 9 tissue slides in each BOND slide tray, HS-HER2 QIF staining was carried out by the following protocol; deparaffinization with BOND dewax solution (AR9222), antigen retrieval with BOND HIER epitope retrieval solution 2 (AR9640) at 97°C for 20 minutes, blocking with ReadyProbes Endogenous HRP & AP blocking solution (R37629, Invitrogen) for 10 minutes and with BSA for 30 minutes, 1 hour-incubation with primary rabbit monoclonal HER2 antibody (clone 29D8, 2165, IgG, Cell Signaling) at optimal concentration of 1ug/ml mixed together with 1:100 concentration of pan-CK (Clones AE1/AE3,M3515, Dako), amplification with Rabbit Envision+ System – HRP labelled polymer anti-Rabbit (K400311-2, Dako) mixed together with 1:100 dilution of a green-fluorescent Alexa Fluor 546 Goat-anti-Mouse IgG (H+L)Cross-Adsorbed Secondary Antibody (A11003, Invitrogen) for 1 hour, staining with 1:50 dilution of a red-fluorescent Tyramide Signal Amplification (TSA) Cyanin 5 (SAT705A001EA, Akoya Biosciences) for 10 minutes and nuclear staining with 1:500 dilution of a blue-fluorescent 4′,6-diamidino-2-phenylindole (DAPI) for 10 minutes.

Techniques: Comparison

Journal: medRxiv

Article Title: Validation and Prospective Testing of a High Sensitivity, Quantitative Analytic Assay for HER2 on Histopathology Slides

doi: 10.1101/2025.05.06.25327097

Figure Lengend Snippet: A) Box plot showing the HER2 proteins in IHC subgroups quantified by HS-HER2 assay; B) HER2 measurement (amol/mm 2 ) in different subgroups stratified by traditional IHC scores (left) and by HS-HER2 assay limits (right); C) Distribution of HS-HER2 measurements split by hormone receptor status

Article Snippet: By loading one control CMA slide together with every 9 tissue slides in each BOND slide tray, HS-HER2 QIF staining was carried out by the following protocol; deparaffinization with BOND dewax solution (AR9222), antigen retrieval with BOND HIER epitope retrieval solution 2 (AR9640) at 97°C for 20 minutes, blocking with ReadyProbes Endogenous HRP & AP blocking solution (R37629, Invitrogen) for 10 minutes and with BSA for 30 minutes, 1 hour-incubation with primary rabbit monoclonal HER2 antibody (clone 29D8, 2165, IgG, Cell Signaling) at optimal concentration of 1ug/ml mixed together with 1:100 concentration of pan-CK (Clones AE1/AE3,M3515, Dako), amplification with Rabbit Envision+ System – HRP labelled polymer anti-Rabbit (K400311-2, Dako) mixed together with 1:100 dilution of a green-fluorescent Alexa Fluor 546 Goat-anti-Mouse IgG (H+L)Cross-Adsorbed Secondary Antibody (A11003, Invitrogen) for 1 hour, staining with 1:50 dilution of a red-fluorescent Tyramide Signal Amplification (TSA) Cyanin 5 (SAT705A001EA, Akoya Biosciences) for 10 minutes and nuclear staining with 1:500 dilution of a blue-fluorescent 4′,6-diamidino-2-phenylindole (DAPI) for 10 minutes.

Techniques:

Journal: bioRxiv

Article Title: SRSF3 is oncogenic in breast but tumor-suppressive in liver by differential regulation of gene expression

doi: 10.1101/2025.03.14.643315

Figure Lengend Snippet: Srsf3-knockout (KO) delays development of Erbb2 breast cancer, but promotes liver cancer induction by DEN. (A) Flaw chart of mouse mammary-specific Srsf3 KO and Erbb2 breast cancer induction and liver-specific Srsf3 KO and liver cancer induction by DEN. Mammary cancer induction was assessed by crossing Srsf3-WT, hetKO (heterozygous knockout), KO female mice with male mice carrying a homozygous V664E mutant of rat c-neu/Erbb2 of which expression is under control by a MMTV promoter (MMTV). Liver cancer induction in mice with liver-specific Srsf3 knockout was performed by the intraperitoneal (i.p.) injection of diethylnitrosamine (DEN) at 25 mg/kg at 15 days of age. Tumor formation was surveyed by palpation weekly up to 18 months of age. (B) Erbb2 (c-neu) staining of Srsf3 WT and Srsf3 KO mammary tissues at six months of age. Six of six Srsf3 WT mice examined developed Erbb2-tumors, but none of all eight (0/8) Srsf3 KO mice had Erbb2 tumors. Scale bar, 2 mm. (C) H&E staining of Srsf3 WT and Srsf3 KO liver tissues at age of six months. Seven of nine (7/9) Srsf3 KO mice developed hepatocellular carcinoma (HCC), but only one of 15 Srsf3 WT mice developed HCC. Black arrow, karyomegaly. Blue arrow, intranuclear intracytoplasmic invagination. Scale bar, 100 µm. The p-values in (B) and (C) were calculated by Chi-squared test. (D, F) Kaplan-Meyer curves for the observed tumor formation rates were plotted till 18 months of age for Erbb2 breast cancer (D) and DEN-induced liver cancer (F) in female mice with or without mammary Srsf3 hetKO or homozygous Srsf3 KO. (E) DEN-induced liver tumor in male mice with or without liver heterozygous or homozygous Srsf3 KO. The p-values were determined by log-rank test between indicated groups.

Article Snippet: Following antibodies are used for protein detection in Western blot and immunohistochemistry: anti-estrogen receptor α (ERα) rabbit polyclonal antibody (MC-20) (Santa Cruz Biotechnology, # sc-542), rabbit polyclonal anti-SRSF3 antibody (Abcam, # ab125124), anti-SRSF1 mouse monoclonal antibody (clone 96) (Thermo Fisher scientific, # 32-4500), anti-HER2/ErbB2 rabbit monoclonal antibody (Cell Signaling Technology, # 2165) for mouse and rat c-neu/Erbb2, anti-Eif4a2 rabbit polyclonal antibody (Abcam, # ab31218), anti-β-tubulin antibody (Sigma, # T5201), anti-β-actin mouse monoclonal antibody (AC-15) (Santa Cruz Biotechnology, # sc-69879), and anti-GAPDH monoclonal antibody (Cell Signaling Technology, # 2118) for GAPDH.

Techniques: Knock-Out, Mutagenesis, Expressing, Control, Injection, Staining