After 2 h of reperfusion, serum samples were isolated from blood samples by centrifugation at 3000 rpm for 10 min at 4°C. The levels of serum cardiac markers CK‐MB, cTnl and the enzyme lactic dehydrogenase (LDH) were measured using commercially available CK‐MB, cTnl and LDH kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China).
Ck mb
Manufactured by Nanjing Jiancheng
Sourced in China
CK-MB is a laboratory test that measures the level of creatine kinase-MB, an enzyme found primarily in the heart muscle. It is used to help detect and evaluate conditions related to heart muscle damage, such as a heart attack.
Automatically generated - may contain errors
Lab products found in correlation
P jnk, by Cell Signaling Technology (2 mentions)
P erk, by Cell Signaling Technology (2 mentions)
Glutathione (gsh), by Nanjing Jiancheng (2 mentions)
Captopril tablets, by Bristol-Myers Squibb (2 mentions)
P nf κb p65, by Cell Signaling Technology (1 mentions)
α tubulin, by Abcam (1 mentions)
Sod test kit, by Nanjing Jiancheng (1 mentions)
Gsdmd, by Abcam (1 mentions)
P iκbα iκbα, by Cell Signaling Technology (1 mentions)
Txnip, by Abcam (1 mentions)
Ck mm, by Nanjing Jiancheng (1 mentions)
A020 2 2, by Nanjing Jiancheng (1 mentions)
Microplate reader, by Agilent Technologies (1 mentions)
Pr6000b, by R&D Systems (1 mentions)
E006 1 1, by Nanjing Jiancheng (1 mentions)
Il 18 elisa kit, by MultiSciences Biotech (1 mentions)
Il 1β elisa kit, by MultiSciences Biotech (1 mentions)
P iκbα, by Cell Signaling Technology (1 mentions)
P ikkβ, by Cell Signaling Technology (1 mentions)
P ikkα, by Cell Signaling Technology (1 mentions)
37 protocols using ck mb
1
Serum Cardiac Marker Analysis
2
Serum Creatine Kinase Isoenzyme Measurement
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The serum was collected, and the content of serum creatine kinase isoenzyme (CK-MB) was detected by the corresponding kit purchased from Nanjing Jiancheng Co., Ltd (Nanjing, Jiangsu, China).
3
Molecular Mechanisms of Inflammation
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The primary antibodies of HSP70, HSP90, TLR4, MyD88, NLRP3, ASC, caspase-1, caspase-11, IL-18, IL-1β, HMGB1, COX-2, iNOS, and caspase-3 were supplied by ABclona Technology (China). Moreover, p-JNK, p-ERK, p-p38, p-IκBα/IκBα, and p-NF-κB (p65) were purchased from Cell Signaling (USA); GSDMD, Bax Trx-1, α-Tubulin, and Txnip were afforded by Abcam (USA). Additionally, CK-MB, CAT, GSH, MDA, and SOD test kits were supplied by Nanjing Jiancheng Biotech. Co., Ltd. (China). Unless specified otherwise, all other reagents were obtained from Sigma–Aldrich (USA).
4
Liver Lipid Profiling in Serum and Tissue
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The serum samples were collected after centrifuging at 3000 rpm, 4 °C for 15 min and stored at − 80 °C. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), creatine kinase MB (CK-MB), creatine kinase MM(CK-MM) concentrations after indicated treatments were determined by the related kits according to the manufacturer’s instructions. The ALT, AST, TG, CK-MB and CK-MM kits were purchased from Nanjing Jiancheng Bioengineering Institute, NanJing, China.
50 mg of liver tissue was homogenized in 500 µL ethanol absolute, then centrifuged at 2500 rpm for 10 min. Triglyceride (TG), total cholesterol(T-CHO)concentrations after indicated treatments were determined by the related kits according to the manufacturer’s instructions.The TG, T-CHO kits were purchased from Nanjing Jiancheng Bioengineering Institute, NanJing, China.Hepatic triglyceride and total cholesterol levels was normalized to tissue wet weight and expressed as mg/g of liver.
50 mg of liver tissue was homogenized in 500 µL ethanol absolute, then centrifuged at 2500 rpm for 10 min. Triglyceride (TG), total cholesterol(T-CHO)concentrations after indicated treatments were determined by the related kits according to the manufacturer’s instructions.The TG, T-CHO kits were purchased from Nanjing Jiancheng Bioengineering Institute, NanJing, China.Hepatic triglyceride and total cholesterol levels was normalized to tissue wet weight and expressed as mg/g of liver.
5
Biomarkers for Cardiac Injury Assessment
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Plasma creatinine kinase-MB (CK-MB; Jiancheng Bioengineering Institute, Nanjing, China) and cardiac troponin I (cTnI; Life Diagnostic, Pennsylvania, USA) from jugular vein blood at various time points (0, 7 and 14day) were determined with ELISA detection kits as per manufacturer's instructions. After treatment for 2 weeks, the serum was collected to detect the concentration of BNP with a BNP ELISA detection kit (BD Biosciences, San Jose, CA) according to the manufacturer's instructions.
6
Quantifying Cardiac Damage Markers
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Blood samples (0.5 mL) were collected from the rats at the end of the study; the sera were separated and stored at −20°C for measuring cardiac damage markers aspartate aminotransferase (AST), creatine kinase-MB (CK-MB), and lactate dehydrogenase (LDH) with commercial kits from Nanjing Jiancheng Bioengineering (Nanjing, China).
7
Enzymatic Biomarker Analysis in Serum
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Collected blood samples were centrifuged at 3,000 r/min for 15 min at 4°C and the serum was obtained. The activities of creatine kinase (CK), creatine kinase-MB (CK-MB), and lactate dehydrogenase (LDH) were measured using commercially available kits (Jiancheng Bioengineering, Nanjing, China) based on the manufacturer’s instruction.
8
Cardiac Biomarker Evaluation Protocol
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Serum lactate dehydrogenase (LDH), creatine kinase MB (CK-MB), and aspartate transaminase (AST) activities were measured using commercial kits (Nanjing Jiancheng Bioengineering, Nanjing, China) based on the provided directions. In addition, serum cTn-I levels were measured via ELISA.
9
Oxidative Stress Biomarkers in Rat Carotid Artery
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Right carotid artery blood samples (2 mL) of rats in each group were taken at T0 and
T5, left standing for 2 h, and centrifuged at 680 g at room
temperature for 10 min. The upper layer of serum was stored at –80°C. Thiobarbituric
acid reaction method was used for the determination of malondialdehyde (MDA),
xanthine oxidase for super oxide dismutase (SOD) and immunosuppressive method for
creatine phosphokinase-MB (CK-MB). The operations were conducted according to the
MDA, SOD and CK-MB (purchased from Nanjing Jiancheng Bio Co., Ltd., Nanjing, China)
kits’ instructions. Plasma concentration of cardiac troponin-I (cTnI) was determined
at T5. In details, blood samples (2 mL) were placed in a clean test tube with
anticoagulant. The operation steps were in strict accordance with the kit (purchased
from Shanghai Seebiotech Biological Technology Co., Ltd., Shanghai, China)
instructions. Colloidal gold immune chromatography was applied and automatic
immunoassay analyzer was used to conduct measurement.
T5, left standing for 2 h, and centrifuged at 680 g at room
temperature for 10 min. The upper layer of serum was stored at –80°C. Thiobarbituric
acid reaction method was used for the determination of malondialdehyde (MDA),
xanthine oxidase for super oxide dismutase (SOD) and immunosuppressive method for
creatine phosphokinase-MB (CK-MB). The operations were conducted according to the
MDA, SOD and CK-MB (purchased from Nanjing Jiancheng Bio Co., Ltd., Nanjing, China)
kits’ instructions. Plasma concentration of cardiac troponin-I (cTnI) was determined
at T5. In details, blood samples (2 mL) were placed in a clean test tube with
anticoagulant. The operation steps were in strict accordance with the kit (purchased
from Shanghai Seebiotech Biological Technology Co., Ltd., Shanghai, China)
instructions. Colloidal gold immune chromatography was applied and automatic
immunoassay analyzer was used to conduct measurement.
10
Circadian Rhythms in Rat Serum Biomarkers
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This experiment was carried out as follows:1 : Fig. S1. The experimental method of Elisa test is in the Additional file 3 : Experimental methods.
From the rats from part 1 of the experiment, the sera from rats sacrificed only at the single time point (08:00) were used to measure the concentrations of creatine kinase-MB (CK-MB; H197; Nanjing JianCheng, Nanjing, China; 24 rats from each of the CA and CM groups); lactate dehydrogenase (LDH; A020-2–2; Nanjing JianCheng; 30 rats from each of the CA and CM groups); and brain natriuretic peptide (BNP; EIAR-BNP-1; RayBiotech, Norcross, GA, USA; 30 rats from each of the CA and CM groups).
From the rats from part 1 of the experiment, sacrificed at one of two time points (08:00 or 20:00), the sera were measured for concentrations of angiotensin II (ANGII; EIAR-ANGII-1; RayBiotech, Norcross, GA, USA; 15 rats from each of the CA and CM groups at each of the 08:00 and 20:00 time points).
Rats from part 2 of the experiment, sacrificed at one of six time points groups (5 rats from each of the CM and CA groups at each time point), the sera were measured for concentrations of epinephrine (EPI, CEA858Ge), norepinephrine (NE, CEA907Ge), and glucocorticoid (COR, CEA462Ge). These three ELISA kits were all obtained from Cloud-Clone Corp. (Wuhan, China).
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