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This immunoassay kit allows for the in vitro quantitative determination of bovine CPTI concentrations in cell culture supernates, serum, plasma and other biological fluids.

Bovine carnitine palmitoyltransferase I [CPT I] ELISA Kit


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Background

Carnitine palmitoyltransferase ? (CPT?) is a mitochondrial enzyme. In muscle and other non-liver tissues, CPT1 is associated with the outer mitochondrial membrane. CPTI mediates the transport of long chain fatty acids across the membrane by binding them to carnitine. This enzyme can be inhibited by malonyl CoA.

Feature and Advantages

<PRINCIPLE OF THE ASSAY>
The microtiter plate provided in this kit has been pre-coated with an antibody specific to CPT?. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for CPT? and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3'5, 5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain CPT?, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of CPT? in the samples is then determined by comparing the O.D. of the samples to the standard curve.

< SPECIFICITY>
This assay recognizes recombinant and natural bovine CPT?. No significant cross-reactivity or interference was observed.

<DETECTION RANGE>
1.25 ng/ml-80 ng/ml.
The standard curve concentrations used for the ELISA’s were 80 ng/ml, 40 ng/ml, 20 ng/ml, 10 ng/ml, 5 ng/ml, 2.5 ng/ml, 1.25 ng/ml.

<SENSITIVITY>
The minimum detectable dose of bovine CPT? is typically less than 0.312 ng/ml.
The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero.

Kit component

  Reagent Quantity
1. Assay plate 1
2. Standard 2
3. Sample Diluent 1 x 20 ml
4. Biotin-antibody Diluent 1 x 10 ml
5. HRP-avidin Diluent 1 x 10 ml
6. Biotin-antibodyt 1 x 120μl
7. HRP-avidin 1 x 120μl
8. Wash Buffer 1 x 20 ml (25×concentrate)
9. TMB Substrate 1 x 10 ml
10. Stop Solution 1 x 10 ml
Product List

Product Name Cat# Quantity Price

Species : 1.25 ng/ml - 80 ng/ml
Assay range : 100 µL (Culture Supernatant, Plasma, Serum)
Assay sample volume : 6-8 hr.+3 hr

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