Sections were prepared from the abovementioned resected specimens (4 μm). Immunohistochemical (IHC) staining for LRG was performed using a rabbit anti‐LRG monoclonal antibody (1:250, ab178698">ab178698 ; Abcam, Chicago, IL, USA), a rabbit anti‐Smad4 monoclonal antibody (1:200, ab40759; Abcam), a rabbit anti‐Smad2 polyclonal antibody (1:100, ab53100; Abcam), a mouse anti‐E‐cadherin polyclonal antibody (610181, 1:200; GE Healthcare Biosciences, Piscataway, NJ, USA) and a mouse anti‐vimentin monoclonal antibody (V6630, 1:200; Sigma‐Aldrich, St. Louis, MO, USA) overnight at 4°C, with visualization using Envision ChemMate (Dako, Glostrup, Denmark), according to the manufacturer's protocol. Three independent gastroenterological oncologists (HW, SK and TO), who were blinded to the histologic data, analyzed the stained sections, which were also photographed using a light microscope (DM2500 with the Leica Application Sweat software program [version 3.80]; Leica Microsystems GmbH, Wetzlar, Germany).
Chemmate envision
Manufactured by Agilent Technologies
Sourced in Denmark
The ChemMate EnVision is a laboratory instrument designed for automated chemical analysis and measurements. It is capable of performing various types of spectroscopic and photometric measurements on liquid samples. The core function of the ChemMate EnVision is to provide accurate and reliable data for analytical applications.
Automatically generated - may contain errors
Lab products found in correlation
Automated stainer, by Agilent Technologies (5 mentions)
Stable dab solution, by Thermo Fisher Scientific (2 mentions)
Ab53100, by Abcam (1 mentions)
Ab178698, by Abcam (1 mentions)
Ab40759, by Abcam (1 mentions)
V6630, by Merck Group (1 mentions)
Anti gpc 1 antibody, by Atlas Antibodies (1 mentions)
Bh 2 microscope, by Olympus (1 mentions)
Vectastain abc rabbit igg kit, by Vector Laboratories (1 mentions)
Hpa001888, by Atlas Antibodies (1 mentions)
Blocking one reagent, by Nacalai Tesque (1 mentions)
Application suite version 3, by Leica (1 mentions)
Dm2500, by Leica (1 mentions)
Cdx2 88, by BioGenex (1 mentions)
Protein block serum free, by Agilent Technologies (1 mentions)
3 3 diaminobenzidine (dab), by Agilent Technologies (1 mentions)
P c met, by Immuno-Biological Laboratories (1 mentions)
C met, by Santa Cruz Biotechnology (1 mentions)
Anti mlh1 g168 728, by BD (1 mentions)
A16 4, by BD (1 mentions)
18 protocols using chemmate envision
1
Immunohistochemical Analysis of Gastric Cancer
2
Immunohistochemical Scoring of GPC-1 in Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Three-micrometre sections were prepared from formalin-fixed, paraffin-embedded tissue samples. As described previously,19 (link) the sections were deparaffinised with xylene and rehydrated in four graded alcohol solutions (70%, 80%, 90% and 100%). Immunohistochemical (IHC) staining for GPC-1 was performed using rabbit polyclonal anti-GPC-1 antibody (Atlas Antibodies AB, Stockholm, Sweden, 1:400) and visualised with Envision ChemMate (Dako, Glostrup, Denmark) according to the manufacturer’s protocol.
Immunostaining was scored according to the intensity of the staining: 0, no staining; 1, normal staining; 2, strong staining. The ‘density’ of staining (termed the positivity score) was as follows: 1, indicates less than 50% positivity; 2, indicates more than 50% positivity. The final IHC score was determined by multiplying the intensity score by the positivity score, resulting in a maximum possible score of 4. These data were referred to as the GPC-1 score. Furthermore, we divided patients into two equally balanced groups by score.
Immunostaining was scored according to the intensity of the staining: 0, no staining; 1, normal staining; 2, strong staining. The ‘density’ of staining (termed the positivity score) was as follows: 1, indicates less than 50% positivity; 2, indicates more than 50% positivity. The final IHC score was determined by multiplying the intensity score by the positivity score, resulting in a maximum possible score of 4. These data were referred to as the GPC-1 score. Furthermore, we divided patients into two equally balanced groups by score.
3
Immunohistochemical Analysis of LRG Protein in Placental and Liver Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human tissue 4-μm-thick sections (placenta and cord, fetal liver) were cut from paraffin blocks, then dewaxed and rehydrated. Rabbit antihuman LRG polyclonal antibody (HPA001888, Atlas Antibodies AB, Stockholm, Sweden, 1:1000) was used as the primary antibody and samples were visualized using EnVision ChemMate (Dako, Glostrup, Denmark) according to the manufacturer’s protocol. For analyses of mouse sections, dewaxed and rehydrated sections (4 μm) were incubated for 20 min in citrate buffer (10 mM citric acid, pH 6.0) at 95–100°C for antigen retrieval. Sections were then treated with 0.3% H2O2 and blocked using Blocking One reagent (Nacalai Tesque), followed by incubation with rabbit anti-mouse LRG1 polyclonal antibody (R322, Immuno-Biological Laboratories Co. Ltd., Gunma, Japan, 1:1,000) overnight at 4°C. After washing, sections were treated with the VECTASTAIN ABC Rabbit IgG Kit (Vector Laboratories, Burlingame, CA, USA). All sections were counterstained with hematoxylin.
Independent obstetricians (Fig 3B ) [E.K. & Ki.Y. & S.M.], Fig 4C [E.K. & M.E. & Ka.Y.]), who were blinded to the histological data, analyzed the stained sections using an Olympus BH2 microscope for Figs 3B and 4C . The intensity of immunostaining was graded as 1 (weak staining), 2 (moderate), or 3 (strong). Grading was determined as the highest grade identified by sampling of 5–10 random fields for each tissue.
Independent obstetricians (
4
Immunohistochemical Analysis of GPC1 in Pancreatic Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Formalin-fixed, paraffin-embedded tissue sections of 5 µm in size for pancreatic cancer primary and matched metastatic tissue microarray were obtained from Tristar Technology Group (Bethesda, MD, USA). The GPC1 expression in tumor tissues was evaluated using a rabbit polyclonal anti-GPC1 antibody (1:2,000, catalog No. GTX104557; GeneTex, San Antonio, TX, USA) and visualized using Envision ChemMate (Dako, Glostrup, Denmark) according to the manufacturer's instructions. Stained sections were also photographed using phase-contrast light microscopy (DM2500 with Leica Application Suite version 3.80; Leica Microsystems GmbH, Wetzlar, Germany).
5
GPC1 Immunostaining Protocol for Tumor Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Formalin-fixed, paraffin-embedded 4-mm tissue sections were prepared. Evaluation of GPC1 expression in tumor tissues was performed using a rabbit polyclonal anti-GPC1 antibody (GeneTex, catalog No. GTX104557, 1:500) and visualized using Envision ChemMate (Dako) according to the manufacturer's instructions. Three independent pathologists (I. Murakami, S. Tsujii,, and K. Yokota) who were blinded to the clinical data analyzed the stained sections, which were also photographed using phase-contrast light microscopy (DM2500 with Leica Application Sweat version 3.80; Leica Microsystems GmbH). Immunostaining was scored according to the intensity of the staining as follows: 0, no staining; 1, week staining; 2, moderate staining; and 3, strong staining. The staining density was scored as either 1 (<50% positivity) or 2 (≥50% positivity). The final immunohistochemistry (IHC) score was determined by multiplying the intensity score by the positivity score, with in a maximum possible score of 6, and was referred to as the GPC1-stained score.
6
Histochemical and Immunohistochemical Characterization of Gastric Lesions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sections of GFM and GH were subjected to alcian blue/periodic-acid Schiff (AB/PAS) double staining to confirm the presence of gastric foveolar epithelium with AB-negative/PAS-positive apical mucin. Subclassification into GFM and GH were made depending on the absence or presence of the oxyntic glands, respectively. The presence of active inflammation (characterised by intraepithelial neutrophils) and foveolar hyperplasia (defined by papillary growth associated with broad stroma) was assessed.
Immunohistochemistry was performed for the adenocarcinomas and representative GFM and GH lesions. Deparaffinized 4-μm thick sections from each paraffin block were exposed to 0.3% hydrogen peroxide for 15 min to block endogenous peroxidase activity. Antigen retrieval was performed by autoclaving in a 10 mM citrate buffer (pH 6.0) for 10 min. Anti-MUC2 (Ccp58; 1 : 200 dilution; Novocastra, Newcastle upon Tyne, England), anti-MUC5AC (CLH2; 1 : 200 dilution; Novocastra), anti-MUC6 (CLH5; 1 : 100, Novocastra), and anti-CDX2 (CDX2-88; 1 : 100; Bio Genex, San Ramon, CA, USA) were used as the primary antibodies. For staining, we used an automated stainer (Dako, Glostrup, Denmark) according to the vendor's protocol. ChemMate EnVision (Dako) methods were used for detection. The staining results were scored as: 0, <10% positive cells; 1+, 11%–50% positive cells; 2+, >50% positive cells.
Immunohistochemistry was performed for the adenocarcinomas and representative GFM and GH lesions. Deparaffinized 4-μm thick sections from each paraffin block were exposed to 0.3% hydrogen peroxide for 15 min to block endogenous peroxidase activity. Antigen retrieval was performed by autoclaving in a 10 m
7
Immunohistochemical Evaluation of VS38c
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
5 μm sections of formalin-fixed paraffin-embedded tissue were cut and then stained by an indirect immunoperoxidase technique using chemMate Envision (Dako, UK). After blocking endogenous peroxidase, sections were incubated with the monoclonal antibody VS38C (Dako, UK) with antigen unmasking pretreatment. Antigens were detected by incubation with labelled polymer and diaminobenzidine. Sections of myeloma were employed as a positive control. Negative controls consisted of sections which were incubated with normal mouse IgG1 (10 or 20 μg/mL, Sigma-Aldrich, Dorset, UK) substituted for the monoclonal antibody. The VS38c staining was scored as strong or weak relative to the positive control. Where the majority of lesional cells were not stained with VS38c, an assessment of the percentage of VS38-positive cells relative to the total number of (lesional) cells was noted.
8
Immunohistochemical Analysis of MUC5B and TTF-1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Three-micrometer-thick sections were made and deparaffinized in xylene, rehydrated in a descending ethanol series, and then treated with 3% hydrogen peroxide for 10 min. After antigen retrieval by autoclaving in 0.01 mol/L citrate buffer (pH 6.0) with 0.1% Tween 20 at 121°C and blocking with 2% normal swine serum (NSS) for 10 min each, the sections were reacted with non-diluted anti-MUC5B monoclonal antibody (hybridoma supernatant) or 200-times-diluted anti-TTF-1 monoclonal antibody (clone; 8G7G3/1, Dako, Glostrup, Denmark) for 2 h at room temperature (RT). After rinsing in Tris-buffered saline (0.01 M Tris-HCl pH 7.5, 150 mM NaCl) three times for 5 min each, the sections were reacted with ChemMATE ENVISION (Dako) for 30 min at RT. The sections were subsequently visualized with Stable DAB solution (Invitrogen; Carlsbad, CA) and counterstained with Mayer's hematoxylin.
9
Histological and Immunohistochemical Analysis of Tumor Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Histological analysis was performed in accordance with WHO classification (Tan et al, 2012 ). Two pathologists (MY and SS) independently scored each case for tumor border, stromal cellularity, stromal atypia, mitosis, stromal overgrowth and malignant heterologous component (Supplementary Table 1 ). Mitotic counts were quantified per 10 high-power fields with the field size adjusted to 0.196 mm2. Discrepant cases were reviewed together to achieve consensus.
Immunohistochemical analysis was performed on formalin-fixed paraffin-embedded specimens. Antigen retrieval was performed by autoclaving in a 10 mM citrate buffer (pH 6.0) for 10 min. Anti-ATRX (HPA001906; 1:200 dilution; Sigma-Aldrich, St. Louis, MO, USA) and anti-DAXX (HPA008736; 1:250 dilution; Sigma-Aldrich) antibodies were used as the primary antibodies (Heaphy et al, 2011 (link)). For staining, we used an automated stainer (Dako, Glostrup, Denmark), according to the manufacturer's protocol. ChemMate EnVision (Dako) methods were used for detection. Tumors showing uniformly negative staining were regarded as showing loss of expression, using lymphocytes and endothelial cells as internal positive controls.
Immunohistochemical analysis was performed on formalin-fixed paraffin-embedded specimens. Antigen retrieval was performed by autoclaving in a 10 mM citrate buffer (pH 6.0) for 10 min. Anti-ATRX (HPA001906; 1:200 dilution; Sigma-Aldrich, St. Louis, MO, USA) and anti-DAXX (HPA008736; 1:250 dilution; Sigma-Aldrich) antibodies were used as the primary antibodies (Heaphy et al, 2011 (link)). For staining, we used an automated stainer (Dako, Glostrup, Denmark), according to the manufacturer's protocol. ChemMate EnVision (Dako) methods were used for detection. Tumors showing uniformly negative staining were regarded as showing loss of expression, using lymphocytes and endothelial cells as internal positive controls.
10
Immunohistochemical PD-L1 Expression Scoring
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemical staining was performed using PD-L1 rabbit antibody (E1L3N™, CST; dilution 1:200) overnight at 4 °C. Immunoreactivity was detected using the DAKO ChemMateEnVision method according to the manufacturer’s instructions. Two pathologists blinded to patients’ information independently assessed expression of PD-L1. Semi-quantitative H score (H-SCORE) was determined by multiplying the percentage of positively stained cells by an intensity score (0, absent; 1, weak; 2, moderate; and 3, strong) and ranged 0–300.
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!