Haematoxylin and bluing reagent

Manufactured by Roche

Haematoxylin and bluing reagent are laboratory staining solutions used in histology and cytology. Haematoxylin is a basic dye that stains nuclei blue-purple, while the bluing reagent is used to enhance and stabilize the staining. These products are commonly used in the preparation and analysis of biological samples for microscopic examination.

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Lab products found in correlation

Benchmark ultra, by Roche (3 mentions) Anti β catenin, by Merck Group (1 mentions) M3619, by Agilent Technologies (1 mentions) Cell conditioner 1, by Roche (1 mentions) Anti podoplanin, by Agilent Technologies (1 mentions) Ultraview universal dab detection kit, by Roche (1 mentions) Optiview amplification kit, by Roche (1 mentions) Cc1 antigen retrieval solution, by Roche (1 mentions) Optiview dab ihc kit, by Roche (1 mentions) Anti pd l1 clone 22c3, by Merck Group (1 mentions) Ab17147, by Abcam (1 mentions) Ab2415, by Abcam (1 mentions) Cell conditioning 1, by Roche (1 mentions) Ultraview dab detection kit, by Roche (1 mentions)

Close spelling variants of this product

Bluing Reagent (58 citations) Haematoxylin (5 citations) Haematoxylin II and bluing reagent (2 citations)

4 protocols using haematoxylin and bluing reagent

Immunohistochemical Staining of Cell Markers

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Sections were pretreated in CC1‐buffer (Cell Conditioner 1, Ventana Medical Systems, Inc, Tucson, AZ, USA) at 95°C for 36 min (E‐cadherin and β‐catenin) and at 95°C for 64 min (podoplanin and Ki‐67). Antibodies were diluted in Ventana antibody diluent and incubation performed at 36°C for 32 min. For detection, Ultra View or Opti View (for podoplanin) Universal DAB Detection kit using a Bench Mark Ultra (Ventana Medical Systems, Inc, Tucson, AZ, USA) was used. Counterstaining was performed with haematoxylin and bluing reagent (Ventana). The following antibodies were used: anti E‐cadherin (M3612, DAKO), diluted 1:25; anti‐β‐catenin (SIGMA‐Aldrich), diluted 1:1500; and anti‐podoplanin (DAKO, M3619), diluted 1:10. In order to map proliferation of the tumours, an antibody against Ki‐67 (CONFIRM, clone 3090; Ventana; Tucson, AZ, USA) ready diluted was also used.

van der Wal J.E., Sgaramella N., Norberg Spaak L., Zborayova K, & Nylander K. (2019). High podoplanin and low E‐cadherin levels correlate with better prognosis in adenoid cystic carcinoma. Clinical and Experimental Dental Research, 5(4), 350-355.

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Immunohistochemical Analysis of Duodenal CCBE1 and Lymphatics

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Formalin-fixed, paraffin-embedded tissue sections from duodenal biopsies were stained on a BenchMark ULTRA automated instrument (Ventana Medical Systems, Inc.). Heat-induced epitope retrieval was performed using CC1 at 95 °C for 64 min followed by antibody incubation at 36 °C for 16 min. The rabbit polyclonal anti-CCBE1 antibody was used at 1:500 dilution (HPA041374, Atlas Antibody, Sigma-Aldrich). The lymphatic specific mouse monoclonal D2–40 antibody was used at a dilution of 1:50 (Zytomed). D2–40, in normal mesenchymal tissue of the gastrointestinal tract, is highly specific for lymphatic endothelial cells and does not cross react with any other tissue type. Binding of anti-CCBE1 or D2–40 primary antibodies to the antigen was visualized using ultraView Universal DAB Detection Kit, an indirect, biotin-free detection system (Ventana). Subsequently sections were counterstained with Haematoxylin and Bluing Reagent (Ventana).

Jackson C.C., Best L., Lorenzo L., Casanova J.L., Wacker J., Bertz S., Agaimy A, & Harrer T. (2015). A Multiplex Kindred with Hennekam Syndrome due to Homozygosity for a CCBE1 Mutation that does not Prevent Protein Expression. Journal of clinical immunology, 36(1), 19-27.

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Immunohistochemical Evaluation of PD-L1 Expression

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Four µm sections were cut from FFPE tissue blocks. After deparaffinization, antigen retrieval was performed by CC1 antigen retrieval solution (pH 8.0, Ventana Medical systems, Tuczon, AZ, USA) on the Ventana BenchMark Ultra automated slide stainer. Slides were incubated with the primary PD-L1 antibody (anti-PD-L1, clone 22C3, Merck) at a dilution of 1/100 for 32 min, followed by visualization with the OptiView DAB IHC Kit and OptiView Amplification Kit (Ventana Medical systems). The specimens were then counterstained with haematoxylin and bluing reagent (Ventana Medical systems) and coverslipped.
PD-L1 expression was scored in immune cells in the paracortex and in the sinuses of the lymph node, excluding the germinal centers. Similar to IDO1 scoring, the intensity of PD-L1 staining in the paracortex was evaluated according to a four-tiered grading system (Supplementary Figure 3A): no expression (0), weak expression (1+), moderate expression (2+) or strong expression (3+). PD-L1 expression in the paracortex was dichotomized into an PD-L1-low group (0 and 1+) and an PD-L1-high group (2+ and 3+). In the sinuses, PD-L1 staining was scored as ‘low’ or ‘high’ (Supplementary Figure 3B).

Meireson A., Ferdinande L., Haspeslagh M., Hennart B., Allorge D., Ost P., Sundahl N., Spaas M., Demeyer A, & Brochez L. (2021). Clinical Relevance of Serum Kyn/Trp Ratio and Basal and IFNγ-Upregulated IDO1 Expression in Peripheral Monocytes in Early Stage Melanoma. Frontiers in Immunology, 12, 736498.

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Evaluating STAT1 Expression in High-Grade Serous Ovarian Cancer

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Tumour STAT1 expression was evaluated using immunohistochemistry (IHC) on an HGSC tissue microarray (TMA) (TFRI‐COEUR) consisting of 0.6 mm diameter FFPE tumour cores, in duplicate, from a cohort of 734 chemotherapy naïve HGSC cases. This large multicentre TFRI‐COEUR cohort contained 550 chemotherapy naïve HGSC cases that formed our STAT1 second independent validation cohort in addition to duplicates for the 184 cases (designated as CHUM cohort; first independent validation 7) from our previous study. The rabbit anti‐human polyclonal STAT1 primary antibody (1:2000 dilution, Abcam #ab2415) 7 and mouse anti‐human monoclonal CD8 primary antibody (1:25 dilution, Abcam # ab17147) were applied on separate TMA slides using the Ventana automated immunostaining system. In brief, antigen retrieval was carried out with Cell Conditioning 1 (Ventana Medical System Inc.) for 60 min. The slides were incubated with primary STAT1 or CD8 antibody with appropriate negative controls, at 37°C for 60 min. Reactions were carried out using the ultraView DAB detection kit (Ventana Medical System Inc.). Slides were counterstained with haematoxylin and bluing reagent (Ventana Medical System Inc.) for 4 min. The IHC stained TMAs were scanned on Aperio Scanscope and digitally conserved.

Au K.K., Le Page C., Ren R., Meunier L., Clément I., Tyrishkin K., Peterson N., Kendall‐Dupont J., Childs T., Francis J., Graham C.H., Craig A.W., Squire J.A., Mes‐Masson A, & Koti M. (2016). STAT1‐associated intratumoural TH1 immunity predicts chemotherapy resistance in high‐grade serous ovarian cancer. The Journal of Pathology: Clinical Research, 2(4), 259-270.

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