Rat Alanine aminotransferase(ALT) ELISA Kit

Instructions
Code CSB-E13024r
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Target Name glutamic-pyruvate transaminase (alanine aminotransferase)
Alternative Names Gpt ELISA Kit; Aat1 ELISA Kit; Gpt1 ELISA Kit; Alanine aminotransferase 1 ELISA Kit; ALT1 ELISA Kit; EC 2.6.1.2 ELISA Kit; Glutamate pyruvate transaminase 1 ELISA Kit; GPT 1 ELISA Kit; Glutamic--alanine transaminase 1 ELISA Kit; Glutamic--pyruvic transaminase 1 ELISA Kit
Abbreviation GPT
Uniprot No. P25409
Species Rattus norvegicus (Rat)
Sample Types serum, plasma, cell culture supernates, tissue homogenates
Detection Range 3.12 mIU/mL-200 mIU/mL
Sensitivity 0.78 mIU/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Metabolism
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<10%      
Three samples of known concentration were tested in twenty assays to assess.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of rat ALT in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)  
1:1 Average % 86  
Range % 80-92  
1:2 Average % 98  
Range % 91-105  
1:4 Average % 100  
Range % 92-110  
1:8 Average % 94  
Range % 86-98  
Recovery
The recovery of rat ALT spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample Type Average % Recovery Range  
Serum (n=5) 95 89-98  
EDTA plasma (n=4) 96 90-100  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
mIU/ml OD1 OD2 Average Corrected  
200 2.814 2.911 2.863 2.778  
100 2.466 2.355 2.411 2.326  
50 1.659 1.564 1.612 1.527  
25 0.853 0.812 0.833 0.748  
12.5 0.469 0.442 0.456 0.371  
6.25 0.243 0.226 0.235 0.150  
3.12 0.149 0.141 0.145 0.060  
0 0.088 0.082 0.085    
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

Citations

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 Q&A
Q:

Kindly address the below queries:
1. The standard ALT enzyme provided in the kit is from which source ?
2. The Standard ALT enzyme given is active or inactive ?
3. Since, we cannot collect more than 150 µL of blood from mice, we can use only 50 µL serum for ELISA. Will it fall within the detectable range ?

A:
We are glad to inform you the following details:
CSB-E13024r,Rat Alanine aminotransferase(ALT) ELISA Kit
Standard:293 cell-derived recombinant protein
Sorry we did not do the activity analysis on the standard.
As to the sample loading volume, we suggest you refer to the manual/datasheet of these 2 kits. If the sample volume is too little, the target might not be recognized during the assay.
Q:

CSB-E13024r
1. In what units, the final data will be represented ? i.e in enzyme activity (IU/L) or
enzyme levels (ng/mg of protein).
2. The reference articles (Article attached) given in the website has mentioned the result
in U/I. What is this unit and how this unit is derived ?
3. Kindly mention the species in which the primary antibody (used in this kit) was raised.

A:
Thanks for your inquiry.
Pls check our answer:
The kit csb-e13024r kit uses two antibodies, mouse monoclonal antibody pre-coated in the plate and goat polyclonal antibody with biotin label. Both of the antibodies are use mIU/mL as the unit.
The kit is quantitatively detected the content of Alanine aminotransferase(ALT).
As for the published reference, pls check this link: https://www.cusabio.com/manage/index.php?action=class&todo=content&m=248&page=1&year=1&code=CSB-E13024r
Any question, pls feel free to contact me.Pls let me know if you have any further questions. Thank you.

Target Background

Function
(From Uniprot)
Catalyzes the reversible transamination between alanine and 2-oxoglutarate to form pyruvate and glutamate. Participates in cellular nitrogen metabolism and also in liver gluconeogenesis starting with precursors transported from skeletal muscles.
Gene References into Functions
  1. Results indicate that AZD4619 increased alanine aminotransferase (ALT1) protein expression in a dose-dependent manner in human, but not in rat hepatocytes. PMID: 27430334
  2. Data suggest that both Alt1 and Alt2 are induced in liver during gluconeogenesis and in diabetes (Goto-Kakizaki rats); in hepatocytes, both Alt1 and Alt2 are induced by glucocorticoid dexamethasone and are suppressed by insulin. PMID: 25865565
  3. Although ALT and GMT activities and TNF-alpha and IL-6 concentrations were higher in male rats histopathological findings have shown slightly more advanced fibrotic and inflammatory changes in female rats. PMID: 17582939
  4. Report the early effects of short-term dexamethasone administration on hepatic and serum alanine aminotransferase in the rat. PMID: 18850354
  5. Protein levels are significantly elevated and correlate with activity, providing the molecular basis for the elevated total serum enzyme activity in liver toxicity. PMID: 19085960

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Subcellular Location Cytoplasm.
Protein Families Class-I pyridoxal-phosphate-dependent aminotransferase family, Alanine aminotransferase subfamily
Tissue Specificity Liver, heart, skeletal muscle, etc.
Database Links

KEGG: rno:81670

STRING: 10116.ENSRNOP00000044411

UniGene: Rn.6318

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