Immunohistochemical staining was performed using the Bond Polymer Refine Detection kit (Leica Biosystems, Newcastle upon Tyne, UK) in the fully automated BondmaX system (Leica Biosystems) on 4-µm-thick formalin fixed, paraffin-embedded sections cut from each tumor sample. Sections were pretreated using heat-mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1; Leica Biosystems) for 30 min at 99 °C. They were then incubated with the primary antibodies for E-cadherin (clone NCH38, 1:200 dilution; Dako Cytomation, Glostrup, Denmark), N-cadherin (clone 6G11, 1:100 dilution; Dako Cytomation), Snail (Polyclonal, 1:100 dilution; Novus Biologicals, Littleton, CO), Zeb1 (Polyclonal, 1:100 dilution; Santa Cruz Biotechnology), and Zeb2 (Polyclonal, 1:100 dilution; Santa Cruz Biotechnology). The slides were ultimately slightly counterstained with hematoxylin. Appropriate positive and negative controls were used. Only membrane immunoreactions were considered to assess E-cadherin and N-cadherin expression, whereas for Snail, Zeb1, and Zeb2, only the nuclear stain was considered. The percentage of immunostained neoplastic cells was assessed. Representative images of each staining are shown in Figure 1 .
E cadherin clone nch38
Manufactured by Agilent Technologies
Sourced in Denmark, United States
E-cadherin (clone NCH38) is a laboratory equipment product offered by Agilent Technologies. It is a mouse monoclonal antibody that recognizes the E-cadherin protein, which is involved in cell-cell adhesion. The core function of this product is to detect and bind to the E-cadherin protein in various research and diagnostic applications.
Automatically generated - may contain errors
Lab products found in correlation
Bond max system, by Leica (2 mentions)
Bond polymer refine detection kit, by Leica (2 mentions)
Snail, by Novus Biologicals (1 mentions)
Pms2 clone ep51, by Agilent Technologies (1 mentions)
Msh6 clone ep49, by Agilent Technologies (1 mentions)
P53 clone do 7, by Agilent Technologies (1 mentions)
Apo transferrin, by Merck Group (1 mentions)
Cholera toxin, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Insulin, by Merck Group (1 mentions)
Hydrocortisone, by Merck Group (1 mentions)
β actin clone ac 74, by Merck Group (1 mentions)
Parthenolide, by Merck Group (1 mentions)
Her2 clone 4b5, by Roche (1 mentions)
Benchmark ultra, by Roche (1 mentions)
Anti cd3 antibody clone 2gv6, by Roche (1 mentions)
Anti cd68 antibody clone kp1, by Agilent Technologies (1 mentions)
Ki 67 clone mib 1, by Agilent Technologies (1 mentions)
Cd56 clone 1b6, by Leica (1 mentions)
6 protocols using e cadherin clone nch38
1
Immunohistochemical Analysis of EMT Markers
2
Immunohistochemical Evaluation of Colorectal Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IHC was performed using the Bond Polymer Refine Detection kit (Leica Biosystems, Newcastle upon Tyne, UK) in the BOND-MAX system (Leica Biosystems). Four-micrometer-thick FFPE sections were incubated with the following primary antibodies: PMS2 (clone EP51; dilution: 1:50 Dako), MSH6 (clone EP49; dilution: 1:50; Dako), E-cadherin (clone NCH-38; dilution: 1:50; Dako), and p53 (clone DO-7; dilution: 1:150; Dako). IHC slides were jointly evaluated by three pathologists (VA, GP, and MF).
Mismatch Repair defective status was assessed by testing PMS2 and MSH6, and samples were defined as dMMR when one or both proteins resulted negative [15 (link)]. In case of protein loss, the dominant component of the heterodimer (i.e., MLH1 for PMS2 and MSH2 for MSH6; Dako) was tested.
p53 was considered as aberrant in the presence of complete loss or diffuse and strong nuclear immunostaining in dysplastic cells [9 (link), 16 (link)].
E-cadherin expression was considered altered in the presence of complete loss or markedly reduced membranous staining (> 30%), regardless of nuclear/cytoplasmic staining [9 (link)].
Mismatch Repair defective status was assessed by testing PMS2 and MSH6, and samples were defined as dMMR when one or both proteins resulted negative [15 (link)]. In case of protein loss, the dominant component of the heterodimer (i.e., MLH1 for PMS2 and MSH2 for MSH6; Dako) was tested.
p53 was considered as aberrant in the presence of complete loss or diffuse and strong nuclear immunostaining in dysplastic cells [9 (link), 16 (link)].
E-cadherin expression was considered altered in the presence of complete loss or markedly reduced membranous staining (> 30%), regardless of nuclear/cytoplasmic staining [9 (link)].
3
Cell Culture Reagents and Antibodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All reagents used for cell culture were purchased from Gibco BRL Co. (Rockville, MD, USA). Cholera toxin, hydrocortisone, insulin, apotransferrin, T3, dimethyl sulfoxide (DMSO), and parthenolide were obtained from Sigma-Aldrich (Sigma-Aldrich Chemical Co., St. Louis, MO, USA). The following antibodies were purchased from their respective sources: Fascin (clone 55K-2, monoclonal mouse anti-human) and E-cadherin (clone NCH-38, monoclonal mouse anti-human) (Dako, Carpentaria, CA, USA); β-actin (clone AC-74) (Sigma-Aldrich Chemical Co.).
4
Immunohistochemical Analysis of HER2 and E-Cadherin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A formalin-fixed, paraffin-embedded FFPE tumor block was cut into 4-μm-thick sections for hematoxylin and eosin (H&E) immunostaining. Immunohistochemistry was performed by using the antibodies for HER2 (clone 4B5, Ventana Medical System, Tucson, AZ, USA) and E-cadherin (clone NCH-38; Dako, Carpinteria, CA, USA). The E-CAD staining was scored on three scales: negative/weak staining (score 0) when less than 10% of tumor cells with strong IHC intensity (2–3+) or less than 30% of cells with weak intensity (1+) were present; reduced staining (score 1) when 10% to 90% of cells showed a strong IHC intensity or about 30% showed a weak intensity; normal staining (score 2) when more than 90% of cells showed a strong IHC intensity.
5
Tissue Microarray Analysis of Immune Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Formalin‐fixed, paraffin‐embedded tumor specimens from patients who met the inclusion criteria were used for tissue microarray construction. We marked one representative tumor area on the H&E‐stained slides. We arrayed a cylindrical 3‐mm tissue core from the corresponding paraffin block into a recipient block using an automated tissue processor (Tissue Microprocessor KIN‐2; Azumaya). In total, 545 patients had adequate cores available for immunohistochemical analysis.
Subsequently, we obtained 4‐μm sections from tissue microarray blocks and stained them with anti‐RNF128 (clone poly, 1:100; Abcam), anti‐CD3 antibody (clone 2GV6, prediluted; Ventana Medical Systems), anti‐CD4 antibody (clone SP35, prediluted; Ventana Medical Systems), anti‐CD45RO antibody (clone UCHL1, 1:200; Leica), anti‐CD25 antibody (clone 4C9, prediluted; Nichirei), anti‐CD68 antibody (clone KP1, 1:50; Dako), anti‐CD163 antibody (clone MRQ‐26, prediluted; Cell Marque), E‐cadherin (clone NCH‐38, prediluted; Dako), and vimentin (clone V9, 1:200; Dako) using a BenchMark ULTRA automated immunohistochemical slide staining system (Ventana Medical Systems) in accordance with the manufacturer's guidelines. Diaminobenzidine was used as the chromogen, whereas hematoxylin was used as the nuclear counterstain. The positive control tissues were stained alongside the study samples.
Subsequently, we obtained 4‐μm sections from tissue microarray blocks and stained them with anti‐RNF128 (clone poly, 1:100; Abcam), anti‐CD3 antibody (clone 2GV6, prediluted; Ventana Medical Systems), anti‐CD4 antibody (clone SP35, prediluted; Ventana Medical Systems), anti‐CD45RO antibody (clone UCHL1, 1:200; Leica), anti‐CD25 antibody (clone 4C9, prediluted; Nichirei), anti‐CD68 antibody (clone KP1, 1:50; Dako), anti‐CD163 antibody (clone MRQ‐26, prediluted; Cell Marque), E‐cadherin (clone NCH‐38, prediluted; Dako), and vimentin (clone V9, 1:200; Dako) using a BenchMark ULTRA automated immunohistochemical slide staining system (Ventana Medical Systems) in accordance with the manufacturer's guidelines. Diaminobenzidine was used as the chromogen, whereas hematoxylin was used as the nuclear counterstain. The positive control tissues were stained alongside the study samples.
6
Immunohistological Analysis of GCC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In archives of the Department of Pathology, First Faculty of Medicine, Charles University and General University Hospital in Prague and the Department of Pathology, Faculty of Medicine, University of Ostrava and University Hospital in Ostrava, a total of nine cases were found that met the diagnostic criteria for GCC.
Immunohistological examination was carried out using the avidin-biotin complex (ABC) method. The following antibodies were used (working dilutions are given in parentheses): AE1-AE3 clone AE1-AE3 (1:50), Ck20 clone Ks 20.8 (prediluted), Ki-67 clone MIB-1 (1:50), Ck7 clone OU-TL 12/13 (1:50), NSE clone 2F11 (1:50), rabbit anti-human somatostatin polyclonal antibody (1:1000), E-cadherin clone NCH-38 (1:50), beta-catenin clone betacatenin-1 (1:400) -all antibodies produced by DAKO, Glostrup, Denmark; synaptophysin clone 27G12 (1:100), chromogranin A clone 5H7 (1:100), CD56 clone 1B6 (1:50), p53 clone DO-7 (1:400) -all antibodies produced by Novocastra, Newcastle-upon-Tyne, UK; and CEA rabbit polyclonal antibody (1:50) -produced by Biogenex.
Immunohistological examination was carried out using the avidin-biotin complex (ABC) method. The following antibodies were used (working dilutions are given in parentheses): AE1-AE3 clone AE1-AE3 (1:50), Ck20 clone Ks 20.8 (prediluted), Ki-67 clone MIB-1 (1:50), Ck7 clone OU-TL 12/13 (1:50), NSE clone 2F11 (1:50), rabbit anti-human somatostatin polyclonal antibody (1:1000), E-cadherin clone NCH-38 (1:50), beta-catenin clone betacatenin-1 (1:400) -all antibodies produced by DAKO, Glostrup, Denmark; synaptophysin clone 27G12 (1:100), chromogranin A clone 5H7 (1:100), CD56 clone 1B6 (1:50), p53 clone DO-7 (1:400) -all antibodies produced by Novocastra, Newcastle-upon-Tyne, UK; and CEA rabbit polyclonal antibody (1:50) -produced by Biogenex.
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!