Human malonyl coenzyme A ELISA Kit

Instructions
Code CSB-E13877h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human Malonyl-coenzyme-A (Malonyl CoA) ELISA Kit is designed to measure Malonyl CoA biological samples, including serum, plasma, and tissue homogenates, in a quantitative fashion. This kit has been verified by multiple tests with advantages of high sensitivity, strong specificity, precision less than 10%, high recovery, and lot-to-lot consistency. Based on the Sandwich-ELISA technique in combination with the enzyme-substrate chromogenic reaction as well as colorimetric measurement, the levels of Malonyl CoA in the sample can be calculated.

Malonyl CoA is located at a central regulatory node in mammalian metabolism to coordinate the fatty acid biochemical metabolism. It is the rate-determining intermediate in fatty acid synthesis but is also an allosteric inhibitor of the rate-setting step in mitochondrial long-chain fatty acid oxidation. Acetyl coenzyme A (acetyl-CoA) carboxylase (ACCase) is responsible for the carboxylation of acetyl-CoA to form malonyl-CoA.

Target Name malonyl coenzyme A
Abbreviation malonyl CoA
Species Homo sapiens (Human)
Sample Types serum, plasma, tissue homogenates
Detection Range 0.625 ng/mL-40 ng/mL
Sensitivity 0.156 ng/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 malonyl CoA in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
 SampleSerum(n=4)
1:100Average %86
Range %80-92
1:200Average %96
Range %92-100
1:400Average %100
Range %93-110
1:800Average %91
Range %85-100
Recovery
The recovery of human malonyl CoA 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) 9083-95
EDTA plasma (n=4)9590-100
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
ng/mllOD1OD2AverageCorrected
402.439 2.252 2.346 2.241
201.857 1.760 1.809 1.704
101.340 1.304 1.322 1.217
50.903 0.890 0.897 0.792
2.50.515 0.504 0.510 0.405
1.250.342 0.319 0.331 0.226
0.6250.213 0.203 0.208 0.103
00.109 0.101 0.105  
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human Malonyl CoA 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.
  • Biotin-labeled Malonyl CoA antibody (100 x concentrate) 1 x 120 μl ---Act as the detection antibody.
  • HRP-avidin (100 x concentrate) 1 x 120 μl --- Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • Biotin-antibody Diluent 1 x 15 ml ---Dilute the high concentration Biotin-antibody to an appropriate working solution.
  • HRP-avidin Diluent 1 x 15 ml ---Dilute the high concentration HRP-avidin solution to an appropriate solution.
  • Sample Diluent 1 x 50 ml---Dilute the sample to an appropriate concentration.
  • Wash Buffer (25 x concentrate) 1 x 20 ml --- Wash away unbound or free substances.
  • TMB Substrate 1 x 10 ml --- Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • Stop Solution 1 x 10 ml --- 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 that 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

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 Q&A
Q:

May I know what's the standard in the kit E13877h, recombinant protein?

A:
Thanks for your inquiry.
CSB-E13877h
Standard is small molecule polymer. Pls let me know if you have any further questions. Thank you.

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