Animal experiments were performed as previously described [22 (link)]. Expressions of SDC2 protein were measured using immunochemical analysis on 5-mm slices of formalin fixed paraffin-embedded tumor xenografts in nude mice. Antigens were retrieved by pretreating dewaxed sections in a microwave oven at 750 W for 5 min in a citrate buffer (pH 6) processed with the Super Sensitive Link-Labeled Detection System (Biogenex, Menarini, Florence, Italy). The enzymatic activities were developed using 3-amino-9-ethylcarbazole (Dako, Milan, Italy) as a chromogenic substrate. Following counterstaining with Mayer hematoxylin (Invitrogen), slides were mounted in aqueous mounting medium (glycergel, Dako). Pictures were taken using a LEICA DM 4000B microscope, while the relative level of each protein was calculated using LEICA software, percentage of the mock over the chemotherapeutic treated tumors was calculated and plotted.
Super sensitive link labeled detection system
Manufactured by BioGenex
Sourced in Italy
The Super Sensitive Link-Labeled Detection System is a laboratory equipment product designed for sensitive detection applications. It utilizes a link-labeled approach to enhance detection capabilities. The core function of this system is to provide a sensitive and reliable method for target analyte identification and quantification.
Automatically generated - may contain errors
Lab products found in correlation
Glycergel, by Agilent Technologies (4 mentions)
3 amino 9 ethylcarbazole, by Agilent Technologies (3 mentions)
Dm4000b microscope, by Leica (3 mentions)
Mayer hematoxylin, by Thermo Fisher Scientific (2 mentions)
Dm4000b, by Leica (1 mentions)
3 amino 9 ethylcarbazole aec, by Agilent Technologies (1 mentions)
Ab8365, by Abcam (1 mentions)
Mayer s hematoxylin, by Thermo Fisher Scientific (1 mentions)
6 protocols using super sensitive link labeled detection system
1
Measuring SDC2 Protein in Tumor Xenografts
2
Xenograft Model for AGTR1 Expression
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The xenograft model on nude mice was generated and analyzed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals. The analysis was performed as previously reported [29 ]. The expression of AGTR1 protein was measured using immunochemical analysis. Antigens were retrieved by pretreating dewaxed sections and processed with the Super Sensitive Link-Labeled Detection System (Biogenex, Menarini, Florence, Italy). Pictures were taken using a LEICA DM 4000B microscope. The animal study proposal was approved by the Institutional Animal Care and Use Committee (IACUC) of the University of Science and Technology of China. All of the mouse experimental procedures were performed in accordance with the Regulations for the Administration of Affairs Concerning Experimental Animals approved by the State Council of People’s Republic of China.
3
Immunohistochemistry for HIF-2α and CD55
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Clinical samples from the biobank of the Institute of Pathology of the University Hospital of Basel were used for the TMA construction as described in Supplementary Material and methods . Expression of HIF-2α and CD55 was analyzed by immunohistochemistry on 5-mm formalin-fixed paraffin-embedded sections of NB by using an antibody to HIF-2-α (Abcam ab8365, Cambridge, UK,) and to CD55 (HPA024386 Sigma-Aldrich). Antigen was retrieved by pretreating dewaxed sections in a microwave oven at 750 W for 5 min in citrate buffer (pH 6) and processing them with a Super Sensitive Link-Labeled Detection System (Biogenex, Florence, Italy). The enzymatic activity was developed using 3-amino-9-ethylcarbazole (AEC, Dako, Gostrup, Denmark) as achromogenic substrate. Following counterstaining with Mayer's haematoxylin, slides were mounted in aqueous mounting medium (glycergel, Dako, Gostrup, Denmark).
4
Immunochemical Analysis of DLL1 Protein
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The expression of DLL1 protein was measured using immunochemical analysis of 5-mm slices of formalin-fixed paraffin-embedded tumor xenografts from the nude mice. The tissue slides from all six groups were placed on a single slide and simultaneously subjected to the same immunestaining. Antigens were retrieved by pretreating dewaxed sections in a microwave oven at 750 watts for 5 min in citrate buffer (pH 6) and processed with the Super Sensitive Link-Labeled Detection System (Biogenex, Menarini, Florence, Italy). The enzymatic activities were determined using 3-amino-9-ethylcarbazole (Dako, Milan, Italy) as a chromogenic substrate. Following counterstaining with Mayer hematoxylin (Invitrogen), the slides were mounted in aqueous mounting medium (glycergel, Dako). Pictures were taken usinga LEICA DM 4000B microscope. Accordingto the rates of positive cell in each field, we marked the chipsas 0 (pigment free), + (light yellow),++(yellow),+++ (51∼75%), and ++++(brownish yellow) by their color intensity52 (link).
5
Immunohistochemical Analysis of ING5 Protein Expression in Tumor Xenografts
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Animal experiments were performed as previously described [21 (link)]. ING5 protein expression was measured using immunohistochemical analysis on 5-mm slices of formalin-fixed paraffin-embedded tumor xenografts in nude mice. To avoid inter-treatment bias, the tissue slides from all six groups were made on a single slide and subjected to the same immunostaining simultaneously. Antigens were retrieved by pretreating dewaxed sections in a microwave oven at 750 W for 5 min in a citrate buffer (pH 6) processed with the Super Sensitive Link-Labeled Detection System (Biogenex, Menarini, Florence, Italy). The enzymatic activities were developed using 3-amino-9-ethylcarbazole (Dako, Milan, Italy) as a chromogenic substrate. Following counterstaining with Mayer's hematoxylin (Invitrogen), slides were mounted in aqueous mounting medium (glycergel, Dako). Pictures were taken using a LEICA DM 4000B microscope, while the relative level of each protein was calculated using LEICA software. The percentage of the mock- over the chemotherapeutic-treated tumors was calculated and plotted.
6
Nude Mouse Xenograft Model Analysis
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A nude mouse xenograft model was established and analyzed according to the National Institutes of Health Guidelines for the Nursing and Use of Laboratory Animals. The analysis was carried out as previously reported [28 (link)]. The CCND2 and PYR1 protein expressions were detected by immunohistochemistry. The antigen was extracted by pretreatment dewaxing section and handled by the Super Sensitive Link-Labeled Detection System (Biogenex, Italy). The pictures were taken using a LEICA DM 4000B microscope. The animal research proposal was approved by IACUC of Anhui Medical University.
Nude mice were bought from Shanghai Slack Laboratory Animal Co., Ltd., and were sacrificed by euthanasia using CO2 inhalation. After the study, the animals were processed together by the IACUC.
Nude mice were bought from Shanghai Slack Laboratory Animal Co., Ltd., and were sacrificed by euthanasia using CO2 inhalation. After the study, the animals were processed together by the IACUC.
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