Immunohistochemical (IHC) analysis was conducted to study GFAP, Ki-67, EGR1, and CCND1 protein expression in glioma xenografts. Briefly, fresh glioma xenografts were fixed in 4% paraformaldehyde, embedded in paraffin, and cut into 5-um-sections. Then, the sections were immunohistochemically stained using Ki-67 antibody (1:100, proteintech, 27,309–1-AP). EGR1 antibody (dilution 1:100; santa cruz, sc-110×), CCND1 antibody (1:100; Abcam, ab134175). Slides were imaged under a light microscope (Leica).
Ki67 antibody
Manufactured by Proteintech
Sourced in China
The Ki67 antibody is a laboratory reagent used in immunohistochemistry and flow cytometry applications. It recognizes the Ki67 protein, a cellular marker for proliferation. The Ki67 antibody can be used to identify and quantify proliferating cells in various samples.
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Lab products found in correlation
Egr1 antibody, by Abcam (1 mentions)
Cell counting kit 8 cck 8, by Wuhan Servicebio Technology (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
β actin antibody, by Proteintech (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
High glucose dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Phenylmethanesulfonyl fluoride, by Wuhan Servicebio Technology (1 mentions)
Bax antibody, by Proteintech (1 mentions)
Horseradish peroxidase hrp labeled secondary antibody, by Abcam (1 mentions)
5 protocols using ki67 antibody
1
Glioma Xenograft Protein Expression Analysis
2
Fluorescent Labeling and Cell Viability Assay
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DOX and Cyanine 5.5 (Cy5.5, C41H44N2O14S4) were purchased from Yuanye (Shanghai, China). Fluorescein isothiocyanate/propidium iodide (PI)/calcein AM, 3, 3′-dioctadecyloxacarbocyanine perchlorate (DiO), BCA protein assay kits, and RIPA lysis buffer were purchased from Beyotime Biotechnology (Shanghai, China). Phosphate-buffered saline (PBS), penicillin-streptomycin, fetal bovine serum, and high-glucose Dulbecco’s modified Eagle’s medium were purchased from Thermo Fisher Scientific (Waltham, MA, USA). 4’, 6-diamidino-2-phenylindole (DAPI), phalloidin, phenylmethanesulfonyl fluoride, and Cell Counting kit-8 (CCK-8) were purchased from Service Bio (Wuhan, China). Caspase 9 antibody, BAX antibody, Ki67 antibody, CD3 antibody, CD4 antibody, and β actin antibody were purchased from Proteintech Company (Wuhan, China). Gp100 was purchased from Affinity Biosciences (Santa Clara, CA, USA). B16 cell lines were donated by the National Engineering Laboratory for AIDS Vaccines at Jilin University (24 (link)), and the use of these cell lines received approval from the institutional review board for research ethics at the China-Japan Union Hospital of Jilin University.
3
Adipocyte Exosomes Promote Breast Cancer
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BALB/C mice (5–6 weeks) were purchased from the Laboratory Animal Center of the Chinese Academy of Medical Sciences (Beijing, China). All mice were bred and maintained under specific pathogen-free conditions. Animal use and experimental procedures were approved by the Animal Care and Use Committee of the Chinese Academy of Medical Sciences. The mice were divided into four groups: one group received a subcutaneous injection of 2 × 106 MCF-7 cells. The second group received an injection of 2 × 106 MCF-7 cells and 1 × 106 MSC-differentiated adipocytes. The other group received an injection of 2 × 106 MCF-7 cells and 1 × 106 MSC-differentiated adipocytes pretreated with 20 μM GW4869 for 48 h. When MSC-differentiated adipocytes were pretreated with 20 μM GW4869 for 48 h, the isolated adipocyte exosomes were undetectable while, in the control group, exosome yield is about 100 μg/107 MSC-differentiated adipocytes. The last group received 2 × 106 MCF-7 cells and 20 μM GW4869. The tumor volume was measured weekly. The tumor tissues were fixed with 10% PFA. Each group was treated with HE and Ki67 staining. Ki67 antibody was purchased from Proteintech.
4
Quantifying Cell Proliferation in Mouse Tumors
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Mouse tumor sections were stained with Ki‐67 antibody (Proteintech, 1 : 10000) to determine cell proliferation. Positively stained color (brown in each case) was selected for quantification of the relative intensity.
5
Nude Mice Xenograft Tumor Model
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A total of 10 female BALB/c nude mice (aged 4–6 weeks old and weighing 18–20 g) were acquired from the Vital River Laboratory Animal Center (Beijing, China). The nude mice were randomly allocated to two groups (the NC and shASAP1-IT1 groups), of 5 mice per group, based on their body weight, and reared in a specific pathogen-free environment. Next, 2×106 cells were implanted subcutaneously into the nude mice to form tumors. When the maximum diameter of the tumors reached about 1 cm, the nude mice underwent uniform euthanasia, and the tumor tissues were stripped, photographed, and weighed. Formalin-fixed and paraffin-embedded tissue sections were incubated with TGFBR1 primary antibody (dilution 1:200; Cell Signaling Technology), Ki67 antibody (dilution 1:200; Proteintech), and CD34 (dilution 1:200; Abcam) overnight at 4 ℃ and the Horseradish peroxidase (HRP)-labeled secondary antibodies (dilution 1:2,000; Abcam) were then incubated. Animal experiments were performed under a project license (No. 2019033) granted by the ethics committee of the Fourth Hospital of Hebei Medical University, in compliance with national guidelines for the care and use of animals. A protocol was prepared before the study without registration.
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