Mouse malonyl coenzyme A ELISA kit

Instructions
Code CSB-E12896m
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The mouse malonyl coenzyme A (CoA) ELISA kit is suitable for the quantitative determination of malonyl CoA levels in mouse serum, plasma, or tissue homogenates. This kit can detect mouse malonyl CoA concentrations in the range of 0.156 ng/mL-10 ng/mL. Its minimum detectable concentration is as low as 0.039 ng/mL. Besides, it also has been validated in other parameters, including specificity, precision, linearity, and recovery.

This kit contains all reagents required for quantifying malonyl CoA levels. Samples and standards are respectively pipetted into wells pre-coated with the anti-mouse malonyl CoA antibody. After incubation, the biotin-conjugated malonyl CoA antibody is added to wells. After thorough washing, the HRP-avidin conjugate is added to wells. Following a wash to remove any unbound substances, the TMB substrate is added to wells and the solution turns blue. The color reaction is terminated after the addition of the stop solution, and the solution turns from blue to yellow. The color intensity develops in proportion to the amount of malonyl CoA bound in the initial step and is measured by reading the absorption value at 450 nm via a microplate reader.

Malonyl-CoA is the S-malonyl derivative of coenzyme A. It plays a key role in chain elongation in fatty acid biosynthesis and polyketide biosynthesis.

Target Name malonyl coenzyme A
Abbreviation malonyl CoA
Species Mus musculus (Mouse)
Sample Types serum, plasma, tissue homogenates
Detection Range 0.156 ng/mL-10 ng/mL
Sensitivity 0.039 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 mouse malonyl CoA 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)  
1:1000 Average % 90  
Range % 84-996  
1:2000 Average % 95  
Range % 89-99  
1:4000 Average % 99  
Range % 95-105  
1:8000 Average % 98  
Range % 90-106  
Recovery
The recovery of mouse 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 Type Average % Recovery Range  
Serum (n=5) 90 86-96  
EDTA plasma (n=4) 95 87-99  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
ng/ml OD1 OD2 Average Corrected  
10 2.214 2.298 2.256 2.159  
5 1.791 1.765 1.778 1.681  
2.5 1.224 1.268 1.246 1.149  
1.25 0.842 0.812 0.827 0.730  
0.625 0.572 0.547 0.560 0.463  
0.312 0.381 0.365 0.373 0.276  
0.156 0.201 0.209 0.205 0.108  
0 0.096 0.098 0.097    
Materials provided
  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-Mouse malonyl CoA 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 malonyl CoA 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
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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