Lan, rottlerin, dynasore, and Cell Count Reagent SF were purchased from Nacalai Tesque, Inc. (Kyoto, Japan), and a lipid peroxidation (LPO) assay kit (BIOXYTECH® LPO-586TM) was provided by OXIS International, Inc. (Portland, Oregon, USA). Nystatin was obtained from Sigma-Aldrich Japan (Tokyo, Japan). Heat-inactivated fetal bovine serum, penicillin, streptomycin, and Dulbecco’s modified Eagle’s medium/Ham’s F12 (DMEM/F12) were purchased from GIBCO (Tokyo, Japan). Benzalkonium chloride (BAC) and a Bio-Rad protein assay kit were obtained from Kanto Chemical Co., Inc. (Tokyo, Japan) and Bio-Rad Laboratories (Hercules, CA, USA), respectively. Ca test kits, mannitol, cytochalasin D, and isoflurane were obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan), and 0.1% pivalephrine and 0.4% benoxil were purchased from Santen Pharmaceutical Co., Ltd. (Osaka, Japan). MC (type SM-4) and HPCD was supplied by Shin-Etsu Chemical Co., Ltd. (Tokyo, Japan) and Nihon Shokuhin Kako Co., Ltd. (Tokyo, Japan), respectively. SUPER FIX was provided by Kurabo Industries (Osaka, Japan). Calpain Activity Fluorometric Assay Kits were purchased from BioVision Inc. (San Francisco, CA, USA).
Calpain activity fluorometric assay kit
Manufactured by Abcam
Sourced in United States
The Calpain Activity Fluorometric Assay Kit is a laboratory tool used to measure the activity of calpain enzymes. It utilizes a fluorogenic substrate to detect and quantify calpain activity in samples. The kit provides a simple and sensitive method for researchers to study calpain function in various biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Mannitol, by Fujifilm (2 mentions)
Isoflurane, by Fujifilm (2 mentions)
Ca test kits, by Fujifilm (2 mentions)
Pivalephrine, by Santen (2 mentions)
Rottlerin, by Nacalai Tesque (1 mentions)
Dynasore, by Nacalai Tesque (1 mentions)
Nystatin, by Merck Group (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Super fix, by Kurabo (1 mentions)
Cell count reagent sf, by Nacalai Tesque (1 mentions)
Heat inactivated fetal bovine serum, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium ham s f 12 dmem f 12, by Thermo Fisher Scientific (1 mentions)
Benzalkonium chloride bac, by Kanto Chemical (1 mentions)
Cytochalasin d, by Fujifilm (1 mentions)
Protein assay kit, by Bio-Rad (1 mentions)
Benoxil, by Santen (1 mentions)
Methylcellulose type sm 4 mc, by Shin-Etsu Chemical (1 mentions)
Synergy htx multi mode reader, by Agilent Technologies (1 mentions)
Jc 1 mitochondrial membrane potential detection kit, by Beyotime (1 mentions)
17 protocols using calpain activity fluorometric assay kit
1
Comprehensive Reagent Sourcing for Cell-based Assays
2
Cyclodextrin-based Ophthalmic Formulation
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The 2-hydroxypropyl-β-cyclodextrin (HPβCD) was obtained from Nihon Shokuhin Kako Co., Ltd. (Tokyo, Japan). Isoflurane, mannitol, and Ca Test Kits were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Methylcellulose type SM-4 (MC) was supplied by Shin-Etsu Chemical Co., Ltd. (Tokyo, Japan). Lanosterol and the LPO Assay Kit (BIOXYTECH® LPO-586TM) were purchased from Nacalai Tesque, Inc. (Kyoto, Japan) and OXIS International, Inc. (Portland, OR, USA), respectively. The Bio-Rad Protein Assay Kit was provided by Bio-Rad Laboratories (Hercules, CA, USA). Benoxil (0.4%) and pivalephrine (0.1%) were obtained from Santen Pharmaceutical Co., Ltd. (Osaka, Japan) and Kanto Chemical Co., Inc. (Tokyo, Japan), respectively. Calpain Activity Fluorometric Assay Kits were provided by BioVision Inc. (San Francisco, CA, USA). All other chemicals were of the highest purity commercially available.
3
Calpain Activity Assay with Zinc
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Calpain activity was measured using Calpain Activity Fluorometric Assay Kit (BioVision). Calpain and its substrate Ac-LLY-AFC provided by the kit were incubated under various concentrations of zinc in the presence of calcium. The fluorescence of the treated sample was examined in situ using a fluorescent plate reader (model SYNERGY/HTX Multi-Mode reader, BioTek) with excitation at 400 nm and emission at 505 nm.
4
Calpain Activity Assay in Mouse Models
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Endogenous calpain enzymatic activity in HdhQ7/Q111 cells, and R6/2 and YAC128 mouse striatum, and recombinant calpain substrate cleavage in the absence/presence of CHIR99021 was measured using the calpain activity fluorometric assay kit (BioVision, Milpitas, CA, USA) following the manufacturer’s instructions.
5
Fluorometric Assay of Calpain Activity
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The activity of calpain was determined using a commercial Calpain Activity Fluorometric Assay Kit (BioVision, San Francisco, CA) according to the manufacturer's instructions. Briefly, cells were resuspended and counted, and then 1×106 cells were lysed in 100 μl extraction buffer and incubated on ice for 20 min. An equal amount of cell lysate, 100 μg total protein, was diluted in extraction buffer up to 85 μl, then 10 μl 10× reaction buffer and 5 μl calpain substrate were added, and incubated at 37°C for 1 h in the dark. Luminesence was recorded in a luminometer (GloxMax™ 20/20, Promega, Madison, WI) equipped with a 400 nm excitation filter and a 505 nm emission filter.
6
Neuroprotective Efficacy Evaluation
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6-OHDA, Ascorbic acid (AA), baicalein, Niferdipine, Verapamil and MTT were purchased from Sigma-Aldrich(St, Louis, Mo). DAPI (4’, 6-diamidino-2-phenylindole) and Calcium indicator Fluo-4-AM dye, BAPTA were purchased from Thermo Fisher Scientific (Waltham, MA). Calpain activity fluorometric assay kit was purchased from Biovision. Mitochondrial membrane potential detection kit (JC-1) was purchased from Beyotime biotechnology (China). Apoptosis antibody sample kit and ß-actin antibody were purchased from Cell Signaling Technology (Danvers, MA).
7
Calpain and Calcineurin Activity Assays
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Calpain activity was measured using the Calpain Activity Fluorometric Assay Kit from Biovision, U.S.A. according to the manufacturer’s instructions. Cells were cultured at pH 6.0 with or without psalmotoxin-1 (PcTX1) (10 nM) or calpeptin (10 µM) for different times. At each time point, cytosolic proteins without cell membrane contamination and lysosomal proteases were extracted by using extraction buffer. The fluorometric assay was based on the detection of the cleavage of the calpain substrate Ac-LLY-AFC. The fluorescence was sequentially imaged with a confocal imaging system (FV2000, Olympus, Tokyo, Japan), and the mean fluorescence intensity measured at 530 nm was quantitatively analyzed. The Calcineurin Cellular Assay Kit (Ambion, U.S.A.) was used to determine calcineurin phosphatase activity according to the manufacturer’s instructions.
8
Quantifying Calpain Activity in Paclitaxel-Treated Cells
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To measure calpain activity, a Calpain Activity Fluorometric Assay Kit (Biovision, Inc.) was used. Briefly, 2 × 105 Pa16C cells were seeded into 6-well plates and treated with increasing doses of paclitaxel for 48 hr before performing the fluorometric calpain activity assay in accordance with the manufacturer’s instructions.
9
Fluorometric Assay for Calpain Activity
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The calpain activities were measured using calpain activity fluorometric assay kit (BioVision), according to the manufacturer's instructions. Briefly, collected cells with different treatments at different time were extracted with an extraction buffer. The extracted cytosolic proteins without contaminations of cell membrane and lysosome proteases were incubated with the reaction buffer and calpain substrate Ac-LLY-AFC at 37°C for 1 hour in dark. Cleavage of a substrate was quantified in a fluorometer equipped with a 400 nm excitation filter and 505 nm emission filter. The untreated cells were taken as negative control. The activity was expressed as relative fluorescent units (RFU).
10
Calpain Activity and Apoptosis Analysis
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Calpain activity was detected using a Calpain Activity Fluorometric Assay Kit (BioVision, California, USA). The results were read at 400 nm excitation and 505 nm emission after incubation at 37 °C for 1 h in the dark. Cell apoptosis was tested using an Annexin V-FITC/PI Apoptosis Detection Kit (Yeasen). After staining, 1× binding buffer was added for flow cytometry.
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