Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), phosphate-buffered saline (PBS) and dimethyl sulphoxide (DMSO) were provided by Transgen (Beijing, China). A Hoechst 33258 staining kit was provided by Keygen Biotech (Nanjing, China). MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] was obtained from Solarbio (Beijing, China). Antibodies against Bcl-2, Bax, caspase-3, caspase-8, caspase-9 and β-actin were purchased from Abcam (Cambridge, UK), and antibodies against PARP and cytochrome c were purchased from Cell Signaling Technology (Beverly, MA, USA). Horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Cell Signaling Technology and Transgen. An Annexin V-PE/7-AAD apoptosis detection kit was provided by Becton-Dickinson (San Jose, CA, USA). Celastrol was obtained from Nanjing Zelang Medical Technology Co., ltd. (Nanjing, China). Stock solutions of celastrol were prepared by dissolving the celastrol powder in DMSO to a concentration of 1 M, and stored at −20°C. The working concentrations of celastrol were made by diluting the stock solution with the culture medium. The final concentration of DMSO in the medium was <0.5%.
Hoechst 33258 staining kit
Manufactured by Keygen Biotech
Sourced in China
Hoechst 33258 staining kit is a laboratory product used to detect and visualize DNA in biological samples. It contains the fluorescent dye Hoechst 33258, which selectively binds to the minor groove of DNA, allowing for the identification and quantification of nucleic acids.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Transgene (1 mentions)
Annexin 5 pe 7 aad apoptosis detection kit, by BD (1 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by Cell Signaling Technology (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Solarbio (1 mentions)
Annexin 5 fitc propidium iodide apoptosis assay kit, by BD (1 mentions)
Fetal bovine serum (fbs), by Cytiva (1 mentions)
4 protocols using hoechst 33258 staining kit
1
Celastrol-Induced Apoptosis in Cells
2
Toosendanin-Induced Apoptosis in Cancer Cells
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Purified toosendanin (source: root bark and bark of Melia toosendan; molecular formula (MF): C30H38O11; molecular weight (MW): 574.62; purity ≥98%) was purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). RPMI-1640 medium, phosphate-buffered saline (PBS), dimethyl sulfoxide (DMSO), bovine serum albumin (BSA), Cell Counting Kit-8 (CCK-8) and horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies (dilution 1:2,500; cat. no. HS101) were purchased from TransGen Biotech, Inc. (Beijing, China). FBS was obtained from HyClone Laboratories (GE Healthcare, Chicago, IL, USA), while the Annexin V-FITC/Propidium Iodide (PI) Apoptosis Assay kit was obtained from Becton Dickinson (BD Biosciences, San Jose, CA, USA). The Hoechst 33258 staining kit was purchased from Nanjing KeyGen Biotech Co., Ltd. (Nanjing, China). Antibodies against B-cell lymphoma 2 (Bcl-2; dilution 1:2,000; cat. no. ab182858), Bcl-2-associated X protein (Bax; dilution 1:1,000; cat. no. ab32503), cytochrome c (dilution 1:5,000; cat. no. ab133504), caspase-3 (dilution 1:5,000; cat. no. ab32351), caspase-8 (dilution 1:1,000; cat. no. ab108333), caspase-9 (dilution 1:1,000; cat. no. ab32539), poly(ADP-ribose) polymerase (PARP; dilution 1:1,000; cat. no. ab32138) and GAPDH (dilution 1:2,500; cat. no. ab9485) were purchased from Abcam (Cambridge, UK).
3
Hoechst33258 Cell Staining Protocol
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The methods for cell culture and transfection are the same as above. The cells were stained using the Hoechst33258 staining kit (KeyGEN Biotech, Beijing, China) following the manufacturer's instruction.
4
Hoechst 33258 Apoptosis Assay
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Cell apoptosis was determined by Hoechst 33258 staining. Cells transfected with siRNA targeting GLYR1 or the negative control were treated with 5-FU at the IC50 for 48 h. Apoptotic cells were detected using the Hoechst 33258 staining kit (KeyGEN BioTECH) according to the manufacturer’s instructions and visualized by fluorescence microscopy.
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