Mouse Aspartate Aminotransferase (AST) ELISA kit

Instructions
Code CSB-E12649m
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
Have Questions? Leave a Message or Start an on-line Chat

Product Details

Description

The product CSB-E12649m is an in vitro ELISA kit for the quantitative measurement of Aspartate Aminotransferase (AST) in mouse serum, plasma, or cell culture supernates. AST is a pyridoxal phosphate (PLP)-dependent transaminase enzyme found in the liver, heart, skeletal muscle, kidneys, brain, and red blood cells. AST plays an important role in amino acid metabolism because, in the presence of PLP (Vitamin B6), it catalyzes the transfer of the amino group from aspartate or glutamate to the corresponding ketoacid. Serum AST level, serum ALT (alanine transaminase) level, and AST/ALT ratio are commonly measured clinically as biomarkers for liver health. So the AST test is used to check the liver damage.

This kit uses the quantitative sandwich-based enzyme immunoassay technique to measure the amount of mouse AST in the sample. Standards and samples are respectively added to the microplate wells pre-coated with an anti-mouse AST antibody. Biotin-labeled AST antibody, HRP-avidin, TMB substrate are pipped into the microplate in turn. The AST in the samples or standards is sandwiched between pre-coated AST antibody and Biotin-conjugated AST antibody, forming immune complexes. The color renders blue after the addition of TMB substrate. The addition of the stop solution into the wells immediately turns the blue into yellow. The concentration of AST in the samples is directly proportional to OD (450nm). Each manufactured lot of this ELISA kit was quality tested for criteria such as sensitivity, specificity, precision, linearity, and lot-to-lot consistency. See the manual for more information on validation.

Target Name glutamic-oxaloacetic transaminase 2, mitochondrial (aspartate aminotransferase 2)
Alternative Names Got2 ELISA kit; Got-2Aspartate aminotransferase ELISA kit; mitochondrial ELISA kit; mAspAT ELISA kit; EC 2.6.1.1 ELISA kit; EC 2.6.1.7 ELISA kit; Fatty acid-binding protein ELISA kit; FABP-1 ELISA kit; Glutamate oxaloacetate transaminase 2 ELISA kit; Kynurenine aminotransferase 4 ELISA kit; Kynurenine aminotransferase IV ELISA kit; Kynurenine--oxoglutarate transaminase 4 ELISA kit; Kynurenine--oxoglutarate transaminase IV ELISA kit; Plasma membrane-associated fatty acid-binding protein ELISA kit; FABPpm ELISA kit; Transaminase A ELISA kit
Abbreviation GOT2
Uniprot No. P05202
Species Mus musculus (Mouse)
Sample Types serum, plasma, cell culture supernates
Detection Range 1.56 mIU/mL-100 mIU/mL
Sensitivity 0.39 mIU/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Signal Transduction
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 mouse AST 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 % 90  
Range % 85-95  
1:2 Average % 93  
Range % 87-98  
1:4 Average % 91  
Range % 89-94  
1:8 Average % 94  
Range % 90-98  
Recovery
The recovery of mouse AST 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) 96 92-99  
EDTA plasma (n=4) 98 94-103  
             
             
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  
100 2.915 2.898 2.907 2.720  
50 2.475 2.432 2.454 2.267  
25 2.015 2.002 2.009 1.822  
12.5 1.376 1.342 1.359 1.172  
6.25 0.731 0.717 0.724 0.537  
3.12 0.472 0.445 0.459 0.272  
1.56 0.312 0.301 0.307 0.120  
0 0.189 0.185 0.187    
Materials provided
  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-Mouse AST antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled AST antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

Citations

Customer Reviews and Q&A

 Customer Reviews

There are currently no reviews for this product.

Submit a Review here

Earn $30 Amazon Card or 20μL/μg CUSABIO Trial Size Antibody. Details of rewards >>

Target Data

Function Catalyzes the irreversible transamination of the L-tryptophan metabolite L-kynurenine to form kynurenic acid (KA). Plays a key role in amino acid metabolism. Important for metabolite exchange between mitochondria and cytosol. Facilitates cellular uptake of long-chain free fatty acids.
Gene References into Functions
  1. GOT2-mediated ATP synthesis may be a pivotal mechanism underlying the protective effect of febuxostat against neurodegeneration in EA PMID: 29107957
  2. it is quite likely that the down regulation of GOT2 after blast exposure is playing a significant role in mitochondrial dysfunction after blast exposure PMID: 23600763
  3. The role of mAspAT in vivo is to control the levels of endogenous kynurenic acid (KYNA) through modulation of the mAspAt in the mouse brain. PMID: 20977429
  4. KAT IV inhibited by equimolar tryptophan and aspartate, and half-life is 6.5 minutes at 65 degree C. PMID: 20482848
Subcellular Location Mitochondrion matrix, Cell membrane
Protein Families Class-I pyridoxal-phosphate-dependent aminotransferase family
Tissue Specificity Detected in brain (at protein level).
Database Links

KEGG: mmu:14719

STRING: 10090.ENSMUSP00000034097

UniGene: Mm.230169

Newsletters

Get all the latest information on Events, Sales and Offers. Sign up for newsletter today.

© 2007-2020 CUSABIO TECHNOLOGY LLC All rights reserved. 鄂ICP备15011166号-1