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.
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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
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Assay Principle |
quantitative |
Measurement |
Sandwich |
Precision |
Intra-assay Precision (Precision within an assay): CV%<8% |
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Three samples of known concentration were tested twenty times on one plate to assess. |
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Inter-assay Precision (Precision between assays): CV%<10% |
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Three samples of known concentration were tested in twenty assays to assess. |
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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. |
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Sample |
Serum(n=4) |
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1:1 |
Average % |
90 |
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Range % |
85-95 |
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1:2 |
Average % |
93 |
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Range % |
87-98 |
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1:4 |
Average % |
91 |
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Range % |
89-94 |
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1:8 |
Average % |
94 |
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Range % |
90-98 |
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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 |
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Serum (n=5) |
96 |
92-99 |
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EDTA plasma (n=4) |
98 |
94-103 |
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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 |
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100 |
2.915 |
2.898 |
2.907 |
2.720 |
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50 |
2.475 |
2.432 |
2.454 |
2.267 |
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25 |
2.015 |
2.002 |
2.009 |
1.822 |
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12.5 |
1.376 |
1.342 |
1.359 |
1.172 |
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6.25 |
0.731 |
0.717 |
0.724 |
0.537 |
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3.12 |
0.472 |
0.445 |
0.459 |
0.272 |
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1.56 |
0.312 |
0.301 |
0.307 |
0.120 |
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0 |
0.189 |
0.185 |
0.187 |
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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
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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
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Troubleshooting
and FAQs |
ELISA kit FAQs |
Storage |
Store at 2-8°C. Please refer to protocol. |
Lead Time |
3-5 working days |