Diosgenin, RIPA buffer was obtained from Sigma Aldrich (St. Louis, MO, USA). RPMI-1640, DMEM medium, fetal bovine serum (FBS), trypsin, penicillin, and streptomycin were purchased from Gibco (Rockville, MD, USA). PI (propidium iodide), Annexin V-FITC/PI apoptosis detection kit, z-VAD-fmk, MTS, Hoechst 33258, and AO/EB (acridine orange/ethidium bromide) were purchased from Abcam (Cambridge, United Kingdom). 5-Ethynyl-2-deoxyuridine (EdU) was obtained from RiboBio (Guangzhou, China). The primary antibodies against GAPDH, Bax, Bcl-2, Caspase- 3, p21, PARP-1, cytochrome c (CYT C), COX IV, β-catenin and GSK3β were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Other antibodies were from Abcam. Hematoxylin–Eosin Staining Kit was from Solarbio (Beijing, China). The EliVision kit was from Maixin Biotech (Fuzhou, China).
Elivision kit
Manufactured by Maixin Group
Sourced in China
The Elivision kit is a laboratory equipment designed for visual inspection and identification of samples. It provides magnified and illuminated views of specimens for detailed observation.
Automatically generated - may contain errors
Lab products found in correlation
Hematoxylin eosin staining kit, by Solarbio (1 mentions)
5 ethynyl 2 deoxyuridine edu, by RiboBio (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Z vad fmk, by Abcam (1 mentions)
β catenin, by Santa Cruz Biotechnology (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Cytochrome c cyt c, by Santa Cruz Biotechnology (1 mentions)
Gsk 3β, by Santa Cruz Biotechnology (1 mentions)
Hoechst 33258, by Abcam (1 mentions)
Caspase 3, by Santa Cruz Biotechnology (1 mentions)
Parp 1, by Santa Cruz Biotechnology (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Bcl 2, by Santa Cruz Biotechnology (1 mentions)
Trim32 antibody, by Santa Cruz Biotechnology (1 mentions)
Goat serum, by Maixin Group (1 mentions)
P rb antibody, by Cell Signaling Technology (1 mentions)
4 protocols using elivision kit
1
Diosgenin-Induced Apoptosis Mechanisms
2
Immunohistochemical Analysis of TRIM32 Expression
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Immunostaining was performed using the Elivision kit from MaiXin (Fuzhou, China). After antigen retrieval in citrate buffer (pH 6.0) for 2 minutes in an autoclave, 0.3% H2O2 was used for 20 minutes. Sections were incubated with goat serum for 15 minutes. Then, the sections were incubated with TRIM32 antibody at 4°C overnight (1:200 dilution; Santa Cruz Biotechnology Inc., Dallas, TX, USA). Then, HRP conjugated polymers were applied to the sections for 30 minutes. 3,3′-Diaminobenzidine plus kit (MaiXin) was used to develop brown stain. Nuclear and cytoplasmic staining was considered as positive staining. For analysis of staining intensity, five views were examined per slide. The staining intensity was graded as 0 (none), 1 (weak), and 2 (moderate/strong). Percentage was graded as 1: 1%–25%, 2: 26%–50%, 3: 51%–75%, and 4: 76%–100%. Intensity and percentage scores were multiplied to give a final score of 0–8. TRIM32 was designated as low expression (score <4) or high expression (overexpression; score ≥4).
3
Immunohistochemical Analysis of ULBP4 in NPC
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Expression of ULBP4 in NPC tissue samples was assessed on immunohistochemistry (IHC) examination. Paraffin-embedded NPC specimens were sliced into 4-μm thick sections, deparaffinised in xylene and rehydrated through a graded ethanol series. Next, the slides were boiled in a pressure cooker until gas discharging for 2.5 min in citrate buffer (Maixin-bioMVS-0101) (pH 6.0). Endogenous peroxidase was blocked by treatment with 3% hydrogen peroxide (H2O2) for 10 min. Subsequently, sections were incubated with mouse anti-human ULBP4 monoclonal antibody (1:1000, R&D Systems, Inc., Minneapolis, MN, USA) for 1h at 37°C; positive and negative controls were also used. After washing with phosphate-buffered saline (PBS) 3 times, tissues were incubated with horseradish peroxidase-conjugated anti-mouse Ig polymer as a second antibody (Elivision kit, Maixin-Bio, Fuzhou, People's Republic of China) for 30 min at 37°C. Finally, sections were visualized using diaminobenzidine (DAB) staining kit, counter-stained with hematoxylin, and observed under the microscope.
4
Immunohistochemistry of Rab18, p-Rb, and Survivin
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Paraffin tissue sections were made, and immunostaining was performed using Elivision kit from MaiXin (Fuzhou, China). After antigen retrieval in citrate buffer (pH 6.0) for 2 minutes in an autoclave, 0.3% hydrogen peroxide was added for 10 minutes. Sections were incubated with goat serum (ready-to-use; MaiXin) to reduce nonspecific binding. Then, sections were incubated with Rab18 antibody (1:300 dilution; Proteintech, Rosemont, IL, USA), p-Rb antibody (1:200 dilution; Cell Signaling Technology, Danvers, MA, USA), and survivin antibody (1:400 dilution; Cell Signaling Technology) at 4°C overnight. After incubation, horse radish peroxidase (HRP)-conjugated polymers were applied to sections. DAB-Plus kit (MaiXin) was used for staining.
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