Protein expression of ROS1 was detected by immunohistochemical staining in the Korean cohort, and it was not performed in the Singapore cohort due to lack of unstained slides. Tissue microarray sections were stained using the Ventana automated immunostainer BenchMark XT (Ventana Medical Systems, Tucson, AZ). The slides were dried at 60°C for 1 hour and deparaffinized using EZ Prep (Ventana Medical Systems) at 75°C for 4 minutes. Cell conditioning was performed using CC1 solution (Ventana Medical Systems) at 100°C for 8 minutes. ROS1 antibody (rabbit monoclonal, clone D4D6, Cell Signaling Technology, Danvers, MA) was diluted to 1:10, followed by treatment, and incubation at 37°C for 2 hours. Signals were detected using the OptiView DAB IHC Detection Kit (Ventana Medical Systems). Counterstaining was performed using Hematoxylin I (Ventana Medical Systems) for 4 minutes at room temperature.
Hematoxylin 1
Manufactured by Roche
Sourced in United States
Hematoxylin I is a lab equipment product used in histology and cytology. It is a staining agent that binds to nucleic acids, particularly DNA, and is commonly used to stain cell nuclei for visualization under a microscope. The core function of Hematoxylin I is to provide a consistent and reliable way to stain and highlight cellular structures for analysis and identification.
Automatically generated - may contain errors
Lab products found in correlation
Optiview dab ihc detection kit, by Roche (4 mentions)
Cc1 solution, by Roche (2 mentions)
Ez prep, by Roche (2 mentions)
Benchmark ultra, by Roche (2 mentions)
Ultra cell conditioning ultra cc1, by Roche (2 mentions)
Benchmark xt, by Roche (1 mentions)
Ventana benchmark xt, by Roche (1 mentions)
Ventana discovery xt, by Roche (1 mentions)
Iview dab detection kit, by Roche (1 mentions)
H 100, by Santa Cruz Biotechnology (1 mentions)
Bluing reagent, by Roche (1 mentions)
Cell conditioning 1, by Roche (1 mentions)
5 protocols using hematoxylin 1
1
Immunohistochemical Detection of ROS1 Protein
2
Androgen Receptor Immunohistochemistry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After heat-induced antigen retrieval using the target retrieval solution ULTRA Cell Conditioning (ULTRA CC1; Ventana Medical Systems, Tucson, AZ, USA; 950–224 ) tissue microarray slides were stained with a ready to use anti-Androgen Receptor (SP107) rabbit monoclonal primary antibody (Cell Marque, Rocklin, CA, USA; 760-4605). Staining was performed using an automated staining system BenchMark ULTRA (Ventana Medical Systems) in accordance with the manufacturer’s instructions, the following solutions were used: OptiView DAB IHC Detection Kit (760–700), Hematoxylin I (790–2208), Bluing Reagent (760–2037).
3
Immunohistochemical Analysis of FFPE Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Formalin-fixed paraffin-embedded (FFPE) tissues were stained using the Ventana BenchMark XT automated immunostainer (Ventana Medical Systems, Tucson, AZ, USA), according to the manufacturer’s instructions. Thereafter, the slides were dried at 60 °C for 1 h and deparaffinized using EZ Prep (Ventana Medical Systems, Tucson, AZ, USA) at 75 °C for 4 min. Cell conditioning was performed using the CC1 solution (Ventana Medical Systems, Tucson, AZ, USA) at 100 °C for 64 min. Subsequently, the slides were incubated with ID-1, ID-2, ID-3, and ID-4 rabbit polyclonal antibodies (Abcam, Cambridge, UK) and E2A rabbit polyclonal antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted to 1:50 at 37 °C for 32 min. The signals were detected using the OptiView DAB IHC Detection Kit (Ventana Medical Systems, Tucson, AZ, USA). Counterstaining was performed using hematoxylin I (Ventana Medical Systems, Tucson, AZ, USA) for 4 min at room temperature.
4
Immunohistochemical Evaluation of c-MET
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After heat-induced antigen retrieval using the target retrieval solution ULTRA Cell Conditioning (ULTRA CC1; Ventana Medical Systems, Tucson, AZ, USA; 950-224) tissue microarray slides were stained with an anti-total c-MET (SP44) rabbit monoclonal primary antibody (Ventana Medical Systems; 790-443). Staining was performed using an automated staining system BenchMark ULTRA (Ventana Medical Systems) in accordance with the manufacturer's instructions, the following solutions were used: OptiView DAB IHC Detection Kit (760-700), Hematoxylin I (790-2208), Bluing Reagent (760-2037). The arrays were independently scored by two pathologists (S.M.-G. and W.R.) blinded to tissue annotations and patient outcomes. For the immunohistochemical semiquantitative assessment of HGFR expression, the product of the scores of staining intensity and quantity of immunoreactive tumor cells was calculated based on the following scoring system: the intensity ranged from 0 = negative, 1 = low, 2 = medium to 3 = high; the quantity comprised 0 = no expression, 1 < 10% of positive cells, 2 = positivity in 10% to 50%, 3 = positivity in 51% to 80%, and 4 = positivity in more than 80%. The final immunoreactive score (IRS) score (ranging from 0 to 12) is obtained by multiplication of the intensity score and the quantity score.
5
Immunohistochemical Evaluation of PPAR-γ in Prostate Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The representative paraffin blocks from 63 radical prostatectomy specimens and TMAs including 667 PCAs were sectioned into 4 µm slices, deparaffinized, and antigens demasked in EDTA buffer, pH 8.5 (Cell Conditioning 1 solution, Ventana). Immunohistochemical analysis was conducted with the Ventana Discovery XT automated staining system (Ventana Medical Systems, Tucson, AZ, USA) according to manufacturer's instruction. Staining of two PPAR-γ antibodies (C26H12, rabbit monoclonal, 1:400, Cell Signaling Technology, Danvers, MA, USA; H100, rabbit polyclonal, 1:25, Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used according to each manufacturer's instruction. Detection of PPAR-γ was carried using the iVIEW DAB Detection Kit (Ventana). All staining reactions were carried out in Tris buffer, pH 7.4-7.8 (Ventana reaction buffer, Ventana). Finally, slides were counterstained with hematoxylin I, followed by bluing reagent (Ventana). Omitting of primary antibody was used for the negative control and urothelial carcinoma in bladder and thyroidal follicular carcinoma were used for the positive control.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!