LDH release was detected using the LDH activity kit (Beyotime, Shanghai, China) as previously described [18 (link)]. Briefly, 50 μl supernatant from each sample was transferred into a new plate, mixed with 50 μl kit's reaction mixture, and incubated for 10 min at room temperature. Stop solution was then added. The OD values at 490 nm and 630 nm were measured using a microplate reader (Epoch, BioTek, USA).
Ldh activity kit
Manufactured by Beyotime
Sourced in China
The LDH activity kits are laboratory equipment designed to measure the activity of the enzyme lactate dehydrogenase (LDH) in samples. LDH is an important enzyme involved in cellular metabolism, and its activity can be used as an indicator of various physiological and pathological conditions.
Automatically generated - may contain errors
Lab products found in correlation
Microplate reader, by Bio-Rad (2 mentions)
Microplate reader, by Thermo Fisher Scientific (2 mentions)
Primary antibody dilution buffer, by Beyotime (1 mentions)
Jc 1 probe, by Solarbio (1 mentions)
Primescript reverse transcription reagent kit, by Takara Bio (1 mentions)
Mtt assay, by Solarbio (1 mentions)
Sybr premix ex taq, by Takara Bio (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Beyotime (1 mentions)
Trizol reagent, by Beyotime (1 mentions)
Annexin 5 fitc kit, by BD (1 mentions)
Radioimmunoprecipitation assay (ripa), by Beyotime (1 mentions)
Beyoecl plus kit, by Beyotime (1 mentions)
Model 680, by Bio-Rad (1 mentions)
Benchmark plus, by Bio-Rad (1 mentions)
M2128, by Merck Group (1 mentions)
Microplate spectrophotometer, by Agilent Technologies (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Mtt assay, by Beyotime (1 mentions)
Multiskan spectrum microplate spectrophotometer, by Thermo Fisher Scientific (1 mentions)
15 protocols using ldh activity kit
1
Quantifying LDH Activity in Cell Culture
2
Comprehensive Biochemical Assays for Cell Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The lactate dehydrogenase (LDH) activity kit, TRIzol® reagent, BCA protein assay kit, primary antibody dilution buffer, RIPA, PMSF, dihydroethidium and the BeyoECL plus kit were obtained from Beyotime Institute of Biotechnology. The IL-6 ELISA kit (cat. no. EK0411) and TNF-α ELISA kit (cat. no. EK0527) were purchased from Boster Biological Technology. SYBR® Premix Ex Taq™ and the PrimeScript reverse transcription reagent kit were obtained from Takara Biotechnology Co., Ltd. The Annexin V-FITC kit was purchased from BD Biosciences. MTT assay and the JC-1 probe were purchased from Beijing Solarbio Science & Technology Co., Ltd. PF was obtained from Hunan QianJinXiangJiang Pharmaceutical Industry Co., Ltd. D-glucose, D-mannitol, 5% bovine serum albumin (BSA) blocking buffer and the Masson's Trichrome stain kit (cat. no. G1340) were purchased from Beijing Solarbio Science & Technology Co., Ltd. All antibodies used for western blotting are detailed in Table I .
3
Platelet Damage Quantification by LDH
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The degree of platelet damage caused by GPRPP-Y8U@P was determined by measuring the amount of lactate dehydrogenase (LDH) by the LDH activity kit (Beyotime Biotechnology) to the manufacturer's protocol [51 (link)]. Briefly, 600 μL of rat fresh platelet-rich plasma (PRP) from fresh anticoagulant blood was placed on the GPRPP-Y8U@P tube (1 mg in 100 μL PBS), PBS (100 μL), and glass samples (1 mg in 100 μL PBS). All samples were incubated at 37 °C for 60 min. The suspension was obtained via centrifugal, transferred to a 96-well plate, and detected following the manufacturer's protocol.
4
Quantifying Cellular LDH Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were assessed for LDH activity using LDH activity kit (C0016; Beyotime Institute of Biotechnology, Haimen, China). The absorbance was measured by spectrophotometry with a microplate reader (model 680; Bio-Rad Laboratories) at 450 nm.
5
Measuring LDH Activity in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Forty-eight hours after transfection, the LDH Activity Kit (Beyotime Institute of Biotechnology, Shanghai) was used to detect the LDH activity of each cell group, according to the manufacturer’s protocol. A microplate reader (Benchmark Plus, Bio-Rad, Hercules, USA) was used to measure absorbance at a wavelength of 450 nm.
6
Quantifying Necroptosis-Induced LDH Release
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LDH release in cell supernatant were measured using LDH activity kit (Beyotime, C0017) according to the manufacturer’s protocol. Relative changes in the release of LDH were presented as fold ± SEM of individual control groups without treatment of necroptosis inducer TSZ and GDCA.
7
MTT and LDH Assays for Evaluating Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell viability of chemical inhibitors and siRNA were determined with a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) (Sigma-Aldrich, M2128) assay. In brief, cells were cultured in each well of 96-well plates with varying concentrations of drugs for 24 h, 48 h or 72 h. Following incubation with MTT (0.5 mg/ml) for 4 h, the supernatant of each well was discarded and the insoluble formazan product was dissolved in 100 μl DMSO. The optical density was measured at 570 nm, with a reference wavelength of 630 nm, on a multiscanner autoreader (M450, Bio-rad, USA). The OD570 in control cells was taken as 100% viability. Cell death was assessed by measuring LDH activity released into the culture medium from damaged cells. LDH activity kit (Beyotime, C0016) was used according to the manufacturer's instructions. Unless otherwise specified, the effects of chemical inhibitors or siRNAs on Gyp-L-induced cell death were measured after 24 h treatment.
8
Cytotoxicity Assay of Cell Cultures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells (1×104/well) were seeded in 96-well plates and transfected with Lipofectamine® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.). MTT (20 µl) was added into each well and incubated for 4 h at 37°C. A total of 150 µl isopropanol was added and the cells were incubated at room temperature in the dark for 20 min. The absorbance was measured using a microplate spectrophotometer (Bio-Tek Instruments, Inc., Winooski, VT, USA) at 492 nm.
LDH activity was measured using LDH activity kits (C0016; Beyotime Institute of Biotechnology, Haimen, China) and the absorbance was measured using a microplate spectrophotometer (Bio-Tek Instruments, Inc.) at 450 nm.
LDH activity was measured using LDH activity kits (C0016; Beyotime Institute of Biotechnology, Haimen, China) and the absorbance was measured using a microplate spectrophotometer (Bio-Tek Instruments, Inc.) at 450 nm.
9
Cell Viability and Cytotoxicity Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell viability was analyzed using MTT assays (Beyotime) at 0, 24, 48 and 72 h after transfection. 20 μL of MTT solution was added to each well and cells were incubated for 4 h at 37°C. Supernatants were removed and formazan crystals were dissolved in 150 μL dimethylsulfoxide (DMSO). Absorbance was measured using a Multiskan Spectrum Microplate Spectrophotometer (Thermo Scientific™, USA) at a wavelength of 492 nm.
Cytotoxicity (LDH activity) levels were measured using LDH activity kits (Beyotime). Absorbance was measured using a Multiskan Spectrum Microplate Spectrophotometer (Thermo Scientific™, USA) at a wavelength of 450 nm.
Cytotoxicity (LDH activity) levels were measured using LDH activity kits (Beyotime). Absorbance was measured using a Multiskan Spectrum Microplate Spectrophotometer (Thermo Scientific™, USA) at a wavelength of 450 nm.
10
Cell Proliferation and Cytotoxicity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
At 24, 48, 72 h after transfection, cell proliferation was measured using the MTT assay and incubation was carried out for 4 h at 37 °C. DMSO was added into the cells and then incubated for 20 min. Optical density (OD) value was read using a microplate reader (Bio-Rad Laboratories, Hercules, CA, USA) at 490 nm.
LDH activity was measured using LDH activity kits (Beyotime Biotechnology, Co., Ltd., Shanghai, China). Optical density (OD) value was read using a microplate reader (Bio-Rad Laboratories) at 450 nm.
LDH activity was measured using LDH activity kits (Beyotime Biotechnology, Co., Ltd., Shanghai, China). Optical density (OD) value was read using a microplate reader (Bio-Rad Laboratories) at 450 nm.
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!