Cells were seeded and treated with the indicated drugs for 48 h, detached from the plates with Trypsin Solution without EDTA (Beyotime Biotechnology, Shanghai, China) and washed with cold PBS. The induction of apoptosis was measured by FCM using the Annexin V-PE/7-AAD Apoptosis Detection Kit (Yeasen Biotech, Shanghai, China). And for detection of DNA breakage, a TUNEL assay was performed following the protocol provided by the manufacturer (Yeasen Biotech).
Trypsin solution without edta
Manufactured by Beyotime
Sourced in China
Trypsin Solution without EDTA is a laboratory reagent commonly used for cell detachment and dissociation. It is a proteolytic enzyme that cleaves peptide bonds, facilitating the separation of cells from culture surfaces or other cells. This product does not contain EDTA, which is a common chelating agent used in some trypsin formulations.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin edta, by Thermo Fisher Scientific (3 mentions)
Annexin 5 pe 7 aad apoptosis detection kit, by Yeasen (1 mentions)
Tunel assay, by Yeasen (1 mentions)
Annexin 5 fluorescein isothiocyanate fitc pi apoptosis detection kit, by BD (1 mentions)
Radioimmunoprecipitation assay ripa lysis buffer, by Solarbio (1 mentions)
Phenylmethylsulfonyl fluoride pmsf, by Solarbio (1 mentions)
Anti human cd24 pe, by Thermo Fisher Scientific (1 mentions)
Anti human ki 67, by BioLegend (1 mentions)
Flow cytometry staining buffer, by MultiSciences Biotech (1 mentions)
Flow cytometry, by BD (1 mentions)
Fitc labeled annexin 5 apoptosis detection kit, by BD (1 mentions)
6 protocols using trypsin solution without edta
1
Apoptosis Quantification by FCM and TUNEL
2
Apoptosis Detection in Cell Cultures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CoCl2, 2,7-dichlorodihyfluorescein diacetate (DCFH-DA), Tris, glycine, Tween-20, SDS, acridine orange (AO), propidium iodide (PI), anti-microtubule associated proteins 1A/1B light chain 3B (LC3I/II; cat. no. L7543) were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). Trypsin-EDTA was obtained from Thermo Fisher Scientific, Inc. (Waltham, MA, USA). Trypsin solution without EDTA was purchased from Beyotime Institute of Biotechnology (Hangzhou, China). The Annexin V-fluorescein isothiocyanate (FITC)/PI apoptosis detection kit was purchased from BD Biosciences (San Jose, CA, USA). Primary antibodies were purchased from CST Biological Reagents, Co., Ltd. (Shanghai, China) or Abcam (Cambridge, UK). Secondary antibodies were purchased from OriGene Technologies, Inc. (Beijing, China). All other chemicals were obtained from Sigma-Aldrich (Merck KGaA). Radioimmunoprecipitation assay (RIPA) lysis buffer and phenylmethylsulfonyl fluoride (PMSF) were purchased from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China).
3
Epirubicin Accumulation and Cell Surface Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For epirubicin accumulation assay, the MCF-7/ADR cells were exposed to 2 μg/ml epirubicin, while 1 μg/ml epirubicin was added to MCF-7, MDA-MB-231 and SUM-1315 cells and then after 24h cells were washed with PBS. Then the intracellular epirubicin level was detected by flow cytometry.
Flow cytometry was used to analyze the expression levels of CD44 (Cluster of differentiation 44) and CD24 (Cluster of differentiation 24). Briefly, cells were isolated with Trypsin Solution without EDTA (Beyotime, Shanghai, China) and resuspended. After incubating with anti-human CD44-APC, anti-human CD24-PE (eBioscience, USA) for 30 minutes at room temperature, cells were washed with PBS and evaluated by flow cytometry (BD Biosciences, USA).
Flow cytometry was used to analyze the expression levels of CD44 (Cluster of differentiation 44) and CD24 (Cluster of differentiation 24). Briefly, cells were isolated with Trypsin Solution without EDTA (Beyotime, Shanghai, China) and resuspended. After incubating with anti-human CD44-APC, anti-human CD24-PE (eBioscience, USA) for 30 minutes at room temperature, cells were washed with PBS and evaluated by flow cytometry (BD Biosciences, USA).
4
Quantification of Stem Cell Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The expression levels of Ki-67, CD44 (Cluster of differentiation 44) and CD24 (Cluster of differentiation 24) were analyzed by flow cytometry.
Cells were dissociated using Trypsin Solution without EDTA (Beyotime, Shanghai, China) and resuspended in Flow Cytometry Staining buffer (MULTI SCIENCES, Hangzhou, China). Cells were stained with anti-human Ki-67 (BioLegend, USA) after permeabilization with 70% ethanol at −20 °C for 1 h. Data were compared by the mean of log fluorescence intensity. Cells were incubated with anti-human CD44-APC, anti-human CD24-PE and the APC and PE isotype control antibodies (eBioscience, USA) for 30 min at room temperature. Cells were washed and evaluated by flow cytometry (BD Biosciences, USA) to determinate population distribution.
Cells were dissociated using Trypsin Solution without EDTA (Beyotime, Shanghai, China) and resuspended in Flow Cytometry Staining buffer (MULTI SCIENCES, Hangzhou, China). Cells were stained with anti-human Ki-67 (BioLegend, USA) after permeabilization with 70% ethanol at −20 °C for 1 h. Data were compared by the mean of log fluorescence intensity. Cells were incubated with anti-human CD44-APC, anti-human CD24-PE and the APC and PE isotype control antibodies (eBioscience, USA) for 30 min at room temperature. Cells were washed and evaluated by flow cytometry (BD Biosciences, USA) to determinate population distribution.
5
Annexin V-PE Apoptosis Assay for PANC-1 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Trypsin Solution without EDTA (C0205, Beyotime) was used to dissociate PANC-1 cells. After collection of 1 × 10 [6 (link)] PANC-1, the cells were washed twice with cold PBS and suspended with 400ul Annexin V. Then 5ul Annexin V PE staining solution was added to the suspension. After the cells were incubated for 5–10 min at 2–8 °C under dark conditions, 5-10ul 7-AAD staining solution was added and incubated for 1–3 min. At last, PANC-1 cells was analyzed through flow cytometry (BD Biosciences, USA).
6
Berberine Induces Apoptosis in Renal Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The apoptotic rate of ACHN and A498 treated with various concentrations(0, 20, 50, 100 µM) of berberine for 24 h were detected using the uorescein isothiocyanate (FITC)-labeled Annexin V Apoptosis Detection kit (BD Biosciences, Franklin Lakes, NJ, USA). Brie y, cells were digested with Trypsin Solution without EDTA (Beyotime), then, washed and centrifugated according to the manufacturer's instructions. FITC-labeled annexin V and propidium iodide were used to stain cells for 15 min. The apoptotic death was determined by annexin V/ PI staining. Then the apoptotic rate of cells was measured by Flow Cytometer (BD FACSCelesta, Becton Dickinson USA) using ow cytometry BD FACSDiva 6.1 software.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!