ELISA kits were used to detect the activity of caspase-3 using the caspase-3 colorimetric assay kit (KeyGEN BioTECH) . MCF-7/ ADR cells were seeded into six-well plates at 5 Â 10 5 cells/well. After overnight incubation, they were pretreated with 0, 20, 30 lM DIM for 2 h before exposed to 10 Gy of c-irradiation. Cells were harvested at 48 h after irradiation and washed with ice-cold PBS, then lysed with ice-cold RIPA lysis buffer (KeyGEN BioTECH) with 1 mmol/L PMSF. Protein concentrations were calculated by BCA assay kits (Thermo Fisher SCIENTIFIC, Beijing, China). Caspase-3 activity was measured with the corresponding detection kit according to the manufacturer's instructions.
Caspase 3 colorimetric assay kit
Manufactured by Keygen Biotech
Sourced in China
The Caspase-3 Colorimetric Assay Kit is a laboratory equipment product designed to quantify the activity of the caspase-3 enzyme, a key mediator of apoptosis or programmed cell death. The kit provides a colorimetric-based method for detecting and measuring caspase-3 levels in cell lysates or purified enzyme preparations.
Automatically generated - may contain errors
Lab products found in correlation
Caspase 9 colorimetric assay kit, by Keygen Biotech (10 mentions)
Microplate reader, by Thermo Fisher Scientific (8 mentions)
Dcfh da, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Bca assay kit, by Thermo Fisher Scientific (1 mentions)
Ripa lysis buffer, by Keygen Biotech (1 mentions)
Microplate reader, by Agilent Technologies (1 mentions)
Caspase 3 colorimetric assay kit, by Abcam (1 mentions)
Facscan flow cytometer, by BD (1 mentions)
Annexin 5 fitc apoptosis detection kit, by Beyotime (1 mentions)
Tunel assay kit, by Keygen Biotech (1 mentions)
Facscalibur, by BD (1 mentions)
Model 680, by Bio-Rad (1 mentions)
Bca protein assay kit, by Keygen Biotech (1 mentions)
Ionomycin, by Santa Cruz Biotechnology (1 mentions)
Rhodamine 123, by Merck Group (1 mentions)
Malondialdehyde assay kit, by Beyotime (1 mentions)
Goat anti rabbit igg h l hrp, by Abcam (1 mentions)
Goat anti rabbit igg fitc, by Abcam (1 mentions)
Pifithrin α, by Santa Cruz Biotechnology (1 mentions)
49 protocols using caspase 3 colorimetric assay kit
1
Caspase-3 Activity Assay in MCF-7/ADR Cells
2
Caspase Activity Quantification Protocol
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Activity of caspase-9 or −3 was analysis by spectrophotometry using caspase-9 colorimetric assay kit or caspase-3 colorimetric assay kit (KeyGen, Nanjing, China), and was carried out and presented according to the described protocol. Briefly, 5×106 cells or 100 mg fresh tumor tissues were washed with cold phosphate-buffered saline (PBS) and resuspended in lysis buffer and incubated on ice for 30 min. Then, 50 µl of the cell suspension, 50 µl of reaction buffer, and 5 µl of caspase-3/−9 substrate were mixed, and then incubated at 37°C for 4 h. The absorbance was measured at 405 nm, and BCA protein quantitative analysis was used as the reference to normal each experiment groups.
3
Quantifying Kidney Caspase-3 Activity
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Relative caspase-3 activity in kidney was detected with a caspase-3 colorimetric assay kit (KeyGEN) according to the manufacturer’s instructions. Optical density (OD) was measured at 405 nm with a microplate reader (Bio-Tek).
4
Caspase-3 Activity Assay in U937 Cells
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We initially treated U937 cells (5×106 cells/dish) with 50 nmol/L IDA and/or 50 nmol/L Ara-C for 0 hours and 12 hours. The cells were then collected and lysed on ice for 1 hour. The concentration of protein was detected using the BCA method. A total of 150 μg of protein was detected using the caspase-3 Colorimetric Assay Kit (Nanjing KeyGen Biotech Co., Ltd., Nanjing, People’s Republic of China).
5
Measuring Caspase-3 Activity in Bladder Cancer
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To determine the caspase-3 activity of the two bladder carcinoma cells, the caspase-3 colorimetric assay kit (Keygen) was used following the protocol. The cells were seeded in the 25 cm2 flask, then propagated in PRMI 1640 medium with PKC-α inhibitors, or transfected by the siRNAs against PKC-α and Dicer. And we use the Lysis Buffer in the kit to collect the cells with inhibitors at 0, 8, 16 and 24 hrs; and the cells with transfection was collected after 24 hrs. And protein contents were estimated employing Bradford reagent, equal amount protein were stained with caspase-3 Substrate in 37°C for 4 hrs. Spectrophotometer was used to analyse the caspase-3 activity at 405 nm.
6
Quantifying Chondrocyte Apoptosis Pathways
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The Annexin V-FITC Apoptosis Detection Kit (Beyotime, C1062L) and the Caspase-3 Colorimetric Assay Kit (Keygen Biotech, Nanjing, China) were utilized to determine cell apoptosis. For the Annexin V-FITC Apoptosis Detection Kit, the treated chondrocyte was suspended in the binding buffer containing Annexin V and incubated for 20 min. Afterwards, the chondrocyte was incubated with PI for 15 min. The percentage of chondrocyte apoptosis was examined by a FACScan flow cytometer (Becton Dickinson, USA). In addition, caspase-3 activity was detected using the Caspase-3 Colorimetric Assay Kit (Biovision, K106-25). The absorbance of each sample was measured by a microplate reader (Tecan, F50) at 450 nm.
7
Quantification of Caspase-3 Activity
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Caspase-3 activity in P19CL6 cells was analyzed by means of Caspase-3 Colorimetric Assay kit (KeyGen, Nanjing, China) following the manufacturer’s instructions. In brief, cells were lysed in ice bath for 1 h and vortexed every 20 min. Then the tissue lysates were centrifuged for 10 min at 12,000g at 4 °C. The supernatant were diluted to 50 µl using cell lysis buffer, incubated with 5 µl of substrate at 37 °C for 4 h in dark and a microplate reader (DNM-9602; Beijing Perlong Medical Instrument Ltd, Beijing, China) was used to determine the absorbance of the samples at 405 nm to quantify the caspase-3 activity.
8
Apoptosis Evaluation in Kidney Tissue
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Apoptosis in the kidney tissue was evaluated by enzymatic labeling of DNA strand breaks using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL) assay kit (KeyGEN) according to the manufacturer's instructions. The expression levels of apoptotic genes (Caspase 3, Fas, and Bax) and anti-apoptotic gene (Bcl-2) in the kidney tissues of each group were further detected using real-time qPCR and immunohistochemistry. The activated Caspase3 in kidney tissues was further detected with Caspase-3 colorimetric assay kit (KeyGEN) according to its manufacturer's instructions
9
Evaluating VU0359595 Effects on MM Cells
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To evaluate the effects of VU0359595 on MM cells, we examined apoptotic cell percentage and relative marker caspase-3 levels in U266 and H929 cells after the treatment of bortezomib with or without VU0359595. After incubation for 12 and 24 h, the cells were stained with Annexin V/ propidium iodide according to the manufacturer's instructions (Biosea, Beijing, China). The samples were then immediately analyzed on a flow cytometer (Becton Dickinson FACS Calibur; Franklin Lake, NJ, USA). For caspase-3 assay, cells were collected and lysed with radioimmunoprecipitation assay (RIPA) buffer on ice for 1 h. The activity of caspase-3 was calculated using the caspase-3 colorimetric assay kit (Keygen, Nanjing, China) at OD405 in the treatment group or in the control group.
10
Measuring Chondrocyte Apoptosis
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Total protein was extracted from the articular cartilage of the rats in each group or the cultured chondrocytes. Protein concentrations were determined using the BCA Protein Assay kit (Nanjing KeyGen Biotech Co., Ltd., Nanjing, China). Caspase-3 activity was measured using the Caspase-3 Colorimetric Assay kit (Nanjing KeyGen Biotech Co., Ltd.). The plates were read at 405 nm using a microplate spectrophotometer (Model 680; Bio-Rad, Hercules, CA, USA).
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