To evaluate the ability of distinct RABVs or viral proteins to induce apoptosis in vitro, NA cells were infected with RABVs (GD-SH-01, HEP-Flury, or individual rRABVs) or transfected with different plasmids expressing single viral proteins. At 1 and 2 days post-infection (dpi), cells were harvested and stained with Annexin-V and propidium iodide (PI) using the Annexin V-FITC/PI Kit (BestBio, China). Here, the ratio of apoptotic cells corresponded to the share of Annexin V+/PI- cells and the ratio of death cells corresponded to the share of Annexin V+ cells. Flow cytometry (FCM) was performed using a Beckman FC500 flow cytometer (Beckman Coulter, MA, United States). Data were analyzed with the corresponding CXP analysis software (Beckman Coulter).
Annexin 5 fitc pi kit
Manufactured by BestBio
Sourced in China
The Annexin V-FITC/PI Kit is a laboratory reagent used for the detection of apoptosis and cell viability. It contains Annexin V conjugated with the fluorescent dye FITC and the DNA-binding dye Propidium Iodide (PI). This kit allows for the simultaneous identification of early apoptotic, late apoptotic, and necrotic cells through flow cytometric analysis.
Automatically generated - may contain errors
Lab products found in correlation
Cxp analysis software, by Beckman Coulter (1 mentions)
Beckman fc500 flow cytometer, by Beckman Coulter (1 mentions)
Facsverse flow cytometry, by BD (1 mentions)
Anti notch1, by Abcam (1 mentions)
Cy3 conjugated affinipure goat anti mouse igg, by Proteintech (1 mentions)
Alizarin red s staining kit, by Solarbio (1 mentions)
Sybr green 1 master mix, by Thermo Fisher Scientific (1 mentions)
Anti osterix, by Abcam (1 mentions)
Mtt kit, by Merck Group (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Anti gapdh antibody, by Proteintech (1 mentions)
Anti cd31, by Abcam (1 mentions)
Cyto id green kit, by Enzo Life Sciences (1 mentions)
Gold 3 chloride hydrate haucl4, by Sinopharm (1 mentions)
Sodium citrate c6h5na3o7 2h2o, by Sinopharm (1 mentions)
Beckman fc500, by Beckman Coulter (1 mentions)
Mtt kit, by Dojindo Laboratories (1 mentions)
Facscalibur, by BD (1 mentions)
8 protocols using annexin 5 fitc pi kit
1
Apoptosis Induction by RABV Proteins
2
Apoptosis Quantification in Colorectal Cancer
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After 24‐h transfection, HCT‐116 and SW480 cells were digested with trypsin without EDTA to obtain single‐cell suspensions. Then, cells were subjected to Annexin V‐fluorescein isothiocyanate (FITC) and propidium iodide (PI) using Annexin V‐FITC/PI kit (BestBio). All samples were analyzed with BD FACSVerse flow cytometry (BD Biosciences). Assays were performed at least three times.
3
Ginsenoside Rg1 Biological Assays
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Ginsenoside Rg1 (purity ≥ 98%, molecular structural formula is shown in Supplementary Figure S1 ) was obtained from Shanghai Tubei Biological Co., Ltd. The MTT kit was obtained from Sigma (United States). Annexin V-FITC/PI kit was obtained from BestBio (Shanghai, China). PI kit and RIPA lysis buffer were obtained from Beyotime Biotechnology (Shanghai, China). The BCIP/NBT alkaline phosphatase chromogenic kit and Alizarin Red S staining kit were obtained from Solarbio (Beijing, China). SYBR Green I Master Mix and reverse transcription kit were obtained from Thermo (United States). Anti-VEGF antibody was obtained from Boster (Wuhan, China). Anti-NOG, anti-Notch1, anti-CD31 and anti-Osterix antibodies were obtained from Abcam (United Kingdom). Anti-Emcn antibody was obtained from Abbkine (Wuhan, China). Horseradish enzyme-labeled goat anti-rabbit IgG and horseradish enzyme-labeled goat anti-mouse IgG were obtained from ZSGB-Bio (Beijing, China). Anti-GAPDH antibody, fluorescein (FITC)-conjugated AffiniPure goat anti-rabbit IgG and Cy3-conjugated AffiniPure goat anti-mouse IgG were obtained from Proteintech (United States). Rabbit anti-CD73/PE conjugated antibody, rabbit anti-CD90/PE conjugated antibody, rabbit anti-CD14/FITC conjugated antibody and rabbit anti-CD19/FITC conjugated antibody were obtained from Bioss (Beijing, China).
4
Synthesis and Characterization of PAV Polymers
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Gold(III) chloride hydrate (HAuCl4) and sodium citrate (C6H5Na3O7·2H2O) were purchased from Sinopharm group Co. Ld. Poly(acryloyl-l -valine) (l -PAV) (Mw 4926 Da, polydispersity 1.24), poly(acryloyl-d -valine) (d -PAV) (Mw 4997 Da, polydispersity 1.15) were synthesized and were characterized according our previous works [17 (link), 23 (link)]. High glucose dulbecco’s modified eagle medium (DMEM) and fetal bovine serum (FBS) were obtained from HyClone (USA). Cell counting Kit-8 (CCK8) assay was purchased from DOJINDO Company (Japan). Acridine Orange (AO), 3-methyladenine (3-MA) and primary antibody (Anti-LC3B) (antibody produced in rabbit, L7543) were obtained from Sigma-Aldrich (USA). Annexin V-FITC/PI Kit was bought from BestBio (Shanghai, China). Cyto-ID Green Kit was purchased from ENZO (USA). 2,7-Dichlorodi-hydrofluorescein diacetate (DCFH-DA), radio immunoprecipitation assay (RIPA) lysis buffer penicillin–streptomycin and secondary antibody (Anti-Rabbit IgG (H + L), A0208) were obtained from Beyotime (China). The Milli-Q water was used throughout the experiments.
5
Exosome Formulation Effects on PC9 Apoptosis
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The effect of different exosome formulations on cell apoptosis of PC9 was assessed by using the AnnexinV-FITC/PI kit (Bestbio, Shanghai, China). Cell proliferation-associated ki67 was evaluated by FCM assay using APC antihuman Ki-67 antibody (Biolegend, San Diego, USA). Flow cytometry data were analyzed by Flow Jo software. Cell Counting Kit-8 (CCK-8) assay was also performed to determine the effect of exosomes on cell proliferation. All procedures were according to the manufacturer’s instruction.
6
RABV-Induced Cell Apoptosis Assay
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To investigate the cell apoptosis induced by the RABVs, the pre-seeded NA cells were infected with rescued RABVs at a MOI of 3, the cells were harvested and stained with annexin-V and propidium iodide (PI) using annexin V-FITC/PI kit (Bestbio, Shanghai, China) at 24 hpi following the manufacturer’s instruction. The cell apoptosis was evaluated with a Beckman FC 500 flow cytometry (Beckman Coulter, Brea, CA, USA), and data were analyzed with CXP software.
7
Evaluating Cell Viability and Apoptosis
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Cell viability was measured by MTT kit (Dojindo, Japan). Briefly, we added 0.5 mg/mL MTT solution to each well for 4 h at different points, after transfection of SGC-7901 cells (5 × 103 cells/well). The medium was removed, and DMSO (150 μL) was added to each well. Then, we measured the absorbance at 490 nm, after 10-mintute oscillation. Additionally, the analysis of cell apoptosis was conducted with the Annexin V-FITC/PI kit (BestBio, Shanghai). In short, after treatment, Annexin V-FITC and PI were used to stain the cells. The cell apoptosis was determined by flow cytometry (BD Biosciences, Franklin Lakes, NJ, USA).
8
Apoptosis Assay of BEL-7402/ADM Cells
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The BEL-7402/ADM cells were incubated in groups and treated with AnnexinV-FITC/PI kit (BestBio, Shanghai, China). The cells were collected according to the instructions and then detected by FACS Calibur flow cytometry (Becton-Dickinson, NJ, U.S.A.).
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