ELISA Kit for Alanine Aminotransferase (ALT) Mus musculus (Mouse) Sandwich ELISA

GPT; ALAT; SGPT; AAT1; GPT1; Serum Glutamic Pyruvic Transaminase; Alanine Transaminase; Glutamate pyruvate transaminase 1; Glutamic--alanine transaminase 1

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  • ELISA Kit for Alanine Aminotransferase (ALT) Packages (Simulation)
  • ELISA Kit for Alanine Aminotransferase (ALT) Packages (Simulation)
  • ELISA Kit for Alanine Aminotransferase (ALT) Results demonstration
  • SEA207Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Alanine Aminotransferase (ALT) and the recovery rates were calculated by comparing the measured value to the expected amount of Alanine Aminotransferase (ALT) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 93-102 96
EDTA plasma(n=5) 92-105 97
heparin plasma(n=5) 92-105 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Alanine Aminotransferase (ALT) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Alanine Aminotransferase (ALT) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Alanine Aminotransferase (ALT) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 87-95% 88-102% 87-95% 82-99%
EDTA plasma(n=5) 92-105% 90-97% 82-94% 80-104%
heparin plasma(n=5) 79-94% 87-101% 79-102% 96-104%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Alanine Aminotransferase (ALT)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Alanine Aminotransferase (ALT). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Alanine Aminotransferase (ALT). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Alanine Aminotransferase (ALT), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Alanine Aminotransferase (ALT) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Enhanced antitumor activity by combining an adenovirus harboring ING4 with cisplatin for hepatocarcinoma cellsNature: 201067
  • Additive effects of nicotine and high-fat diet on hepatic steatosis in male mice.PubMed: 23093702
  • Adrenocorticotropic hormone ameliorates acute kidney injury by steroidogenic-dependent and -independent mechanismsPubMed: PMC3612362
  • Pathogenesis of emerging severe fever with thrombocytopenia syndrome virus in C57/BL6 mouse modelPubMed: PMC3382536
  • Protective effect of Panax ginseng against N-acetyl-p-aminophenol-induced hepatotoxicity in ratsAcademicjournals: Source
  • Adrenocorticotropic hormone ameliorates acute kidney injury by steroidogenic-dependent and-independent mechanismsPubmed: 23325074
  • Protective effect of Et-1 receptor antagonist bosentan on paracetamol induced acute liver toxicity in ratsScienceDirect: S0014299914000235
  • Bifidobacterium pseudocatenulatum CECT7765 promotes a TLR2-dependent anti-inflammatory response in intestinal lymphocytes from mice with cirrhosisPubmed:25657013
  • Resolvin E1 and chemerin C15 peptide do not improve rodent non-alcoholic steatohepatitisPubmed:25681756
  • Assessing the Effect of Leptin on Liver Damage in Case of Hepatic Injury Associated with Paracetamol PoisoningPubMed: 26697061
  • The effects of subacute deltamethrin exposure on rat liver: histochemical, immunohistochemical, and biochemical studyxmlui:11446
  • Advanced Studies in Clinical and Experimental Research in GastroenterologyPubmed:26955389
  • Effects of developmental exposure to silver in ionic and nanoparticle form: A study in ratspubmed:27716437
  • Dihydromyricetin protects against liver ischemia/reperfusion induced apoptosis via activation ofFOXO3a-mediated autophagy.pubmed:27793014
  • Adiponectin protects the rats liver against chronic intermittent hypoxia induced injury throughAMP-activated protein kinase pathway.pubmed:27678302
  • Modulation of gut microbiota contributes to curcumin-mediated attenuation of hepatic steatosis in ratspubmed:28341485
  • Antifibrotic Effect of Lactulose on a Methotrexate-Induced Liver Injury Modelpubmed:28912805
  • Alcohol-associated intestinal dysbiosis impairs pulmonary host defense against Klebsiella pneumoniae.pubmed:28604843
  • A critical role of hepatitis B virus polymerase in cirrhosis, hepatocellular carcinoma, and steatosisPubmed:29321963
  • Rolle von CMKLR1 und dessen Liganden bei der nicht-alkoholischen FettlebererkrankungRegensburg:Source
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  • Brg1 promotes liver regeneration after partial hepatectomy via regulation of cell cyclePubmed: 30787318
  • Corticosterone-mediated microglia activation affects dendritic spine plasticity and motor learning functions in minimal hepatic encephalopathyPubmed: 31437533
  • Liraglutide Attenuates Nonalcoholic Fatty Liver Disease by Modulating Gut Microbiota in Rats Administered a High-Fat DietPubmed: 32149099
  • CD47 Deficiency in Mice Exacerbates Chronic Fatty Diet-Induced Steatohepatitis Through Its Role in Regulating Hepatic Inflammation and Lipid MetabolismPubmed: 32158445
  • Biological Safety and Biodistribution of Chitosan NanoparticlesPubmed: 32340313
  • Effects of Tempol in Lipopolysaccharide-Induced Liver Injury
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  • An Experimental Study: Benefits of Digoxin on Hepatotoxicity Induced by Methotrexate Treatment34804155
  • Taraxasterol mitigates Con A-induced hepatitis in mice by suppressing interleukin-2 expression and its signaling in T lymphocytes34848154
  • Water Specific MRI T1 Mapping for Evaluating Liver Inflammation Activity Grades in Rats With Methionine‐Choline‐Deficient Diet‐Induced Nonalcoholic Fatty Liver …Pubmed:35212074
  • Blockade of mIL‐6R alleviated lipopolysaccharide‐induced systemic inflammatory response syndrome by suppressing NF‐κB‐mediated Ccl2 expression and …Pubmed:35548710
  • Dimethyl fumarate ameliorates autoimmune hepatitis in mice by blocking NLRP3 inflammasome activationPubmed:35605433

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