Human Ischemia Modified Albumin,IMA ELISA Kit

Code CSB-E09594h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The human Ischemia Modified Albumin (IMA) ELISA kit is designed for the quantitative detection of human IMA in serum, plasma, and tissue homogenates. It can assay for the analyte in the sample in the range of 3.12 IU/mL to 200 IU/mL. The minimum detectable dose of this kit is typically less than 0.78 IU/mL. It also has high precision, good linearity, and high recovery.

This assay employs the sandwich enzyme immunoassay technique to qualify IMA concentrations.

Samples and standards are respectively pipetted into the micro ELISA plate wells. The biotin-conjugated IMA antibody is subsequently added to wells. The IMA in the samples or the standards is captured by the anti-human IMA antibody pre-coated on the wells and subsequently ligates to the biotin-labeled IMA antibody. Following a thorough washing, HRP-avidin conjugate is added to the wells. The solution appears a blue coloration after the addition of the TMB solution. Finally, the stop solution is added to terminate the color reaction. The solution color turns blue to yellow, and the color intensity develops in proportion to IMA concentrations in the samples and is measured at 450 nm through a microplate reader.

IMA is a modified albumin protein formed by decreased albumin metal-binding capacity in case of acute ischemic stroke. It is a sensitive and early biochemical marker of ischemia.

Target Name Ischemia Modified Albumin,IMA
Abbreviation IMA
Species Homo sapiens (Human)
Sample Types serum, plasma, tissue homogenates
Detection Range 3.12 IU/mL-200 IU/mL
Sensitivity 0.78 IU/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Others
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 human IMA in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
 SampleSerum(n=4)
1:1Average %86
Range %80-90
1:2Average %99
Range %95-102
1:4Average %103
Range %98-107
1:8Average %94
Range %88-98
Recovery
The recovery of human IMA 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 TypeAverage % RecoveryRange
Serum (n=5) 9590-98
EDTA plasma (n=4)9691-99
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
IU/mlOD1OD2AverageCorrected
2002.497 2.467 2.482 2.363
1002.013 1.973 1.993 1.874
501.300 1.288 1.294 1.175
250.687 0.705 0.696 0.577
12.50.442 0.453 0.448 0.329
6.250.283 0.306 0.295 0.176
3.120.198 0.210 0.204 0.085
00.121 0.117 0.119  
Materials provided
  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-Human IMA antibody.
  • 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 IMA 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 incubating.
  • 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°
  • 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
ELISA Data Analysis Watch ELISA data processing video & download Curve Expert if needed
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

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