Beas-2B and NHBE cells were pretreated for 2 h with 1, 5 and 10 μM ALT and then exposed with 5% CSE for 24 h. The cell supernatants (100 μL) of each group were collected, and LDH activity was measured by using a commercial LDH Kit (Keygen, Nanjing, China) according to the manufacturer’s instructions. The absorbance at 450 nm wavelength was measured using a microplate reader (Themo Multiskan MK3, USA).
Ldh kit
Manufactured by Keygen Biotech
Sourced in China
The LDH kit is a laboratory reagent used to measure the activity of the enzyme lactate dehydrogenase (LDH) in biological samples. LDH is an important enzyme involved in cellular metabolism and can be used as a biomarker for various medical conditions.
Automatically generated - may contain errors
Lab products found in correlation
Sodium chloride (nacl), by Xilong (3 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions)
Total no assay kit, by Beyotime (1 mentions)
Js thermo varioskan flash, by Thermo Fisher Scientific (1 mentions)
Sirna, by GenePharma (1 mentions)
Cacl2 2h2o, by Sinopharm (1 mentions)
Potassium chloride (kcl), by Shanghai Lingfeng Chemical Reagent (1 mentions)
6 protocols using ldh kit
1
Cytotoxicity Evaluation of Cigarette Smoke
2
Quantification of Extracellular LDH Concentration
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One hundred microliters of the cell supernatant in each group was collected for the lactate dehydrogenase (LDH) concentration test according to the LDH kit (Keygen, Nanjing, China) instructions. Then, the reaction mixture was added and incubated in the dark for 30 min at room temperature. The LDH concentration was quantified by measuring the absorbance at 490 nm.
3
Modulating Cardiac Fibroblast Response
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miR-122 inhibitor, specific small interfering RNAs (siRNA) for ELA, apelin and GDF15, and corresponding negative control (NC) siRNAs were purchased from GenePharma (Shanghai, China) and transiently transfected into CFs for 48 h at a concentration of 50 nM using Lipofectamine 3000 reagent (Invitrogen, CA, USA). AngII (1 µM) was then added to rat CFs for 24 h. Nitric oxide (NO) and lactate dehydrogenase (LDH) production in CFs were assayed by using Total NO Assay Kit (Beyotime, Shanghai, China) and LDH kit (KGT02448, KeyGEN, China). The absorbance was measured using a full-wavelength multifunctional microplate reader JS-THERMO Varioskan Flash (Thermo, USA).
4
LDH Release Assay for Cytotoxicity
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For LDH release measurement, the cells were grown under NG or HG conditions for eight weeks. After further incubation for 48 h with Exe-4 (10 nM) and/or DDP (25 µM), the supernatants were collected and centrifuged at 12,000 ×g for 30 min. LDH activity (in U/L) was measured in the cleared supernatants with an LDH kit (KeyGEN BioTECH) to confirm the cytotoxic effects of chemotherapeutic agents by reading absorbance at 450 nm (37°C).
5
Rg1 Modulates LPS-Induced Neuronal Damage
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To observe the effect of Rg1 on LPS-induced neuronal damage in HT22 cells, the activity of LDH released to the medium was detected using an LDH kit (Nanjing KeyGen Biotech Co., Ltd.), according to the manufacturer's protocol. The cells were seeded into 96-well plates (5x103 cells per well). When the cells reached ~60-70% confluence, they were treated with LPS or Rg1 (1, 5 or 10 µM), tempol (50 µM) or apocynin (50 µM). Briefly, the supernatant was collected and reacted with nicotinamide adenine dinucleotide (NAD) and lactate solution at 37˚C for 5 min. The absorbance was detected at 490 nm with a Multiskan™ FC microplate reader (Thermo Fisher Scientific, Inc.) to calculate the activity of LDH.
6
Reagent Acquisition for Biochemical Assays
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GAB (CAS 60142-96-3, purity > 98%) was purchased from Aladdin Industrial Corporation. NaCl (CAS 7647-14-5, purity > 99.5%) was purchased from Xilong Scientific Co., Ltd. CaCl2·2H2O (CAS 10035-04-8, purity > 99.0%) was purchased from Sinopharm Chemical Reagent Co., Ltd. MgSO4·7H2O (CAS 10034-99-8, purity > 99.0%), NaHCO3 (CAS 144-55-8, purity > 99.5%) and KCl (CAS 7447-40-7, purity > 99.5%) were purchased from Shanghai Lingfeng Chemical Reagent Co., Ltd. The CAT kit (Cat. no: KGT017), the GSH kit (Cat. no: KGT006), the GST kit (Cat. no: KGT005), the LDH kit (Cat. no: KGT02448), the·OH kit (Cat. no: KGT010) and the Bradford protein quantitation assay kit (Cat. no: KGT801) were purchased from KeyGEN bio TECH Co., Ltd.
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