详细说明

Human PKC/Protein kinase C ELISA Kit 96T

价格：1800
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产品编号 NG-EA463
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  • 产品细节
  • 操作步骤

This Human PKC ELISA kit is intended Laboratory for Research use only and is not for use in diagnostic or therapeutic procedures. The Stop Solution changes the color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of Human PKC in the sample, this Human PKC ELISA Kit includes a set of calibration standards. The calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of Optical Density versus Human PKC concentration. The concentration of Human PKC in the samples is then determined by comparing the O.D. of the samples to the standard curve.


Human PKC/Protein kinase C ELISA Kit 96T
Assay Type
 Solid Phase Sandwich ELISA
Format
 96-well strip plate/48-well strip plate
Assay Length
 2.5 hours
Sample Type undefined Volume
Required Per Well
 Cell Culture Supernates (10 uL), Serum (10uL),Tissue or cell lysate (10uL)
Sensitivity
 0.25 pmol/L
Assay Range
 0.25-8 pmol/L(Cell Culture Supernates, Serum,Tissue or cell lysate)
Specificity
 Natural and recombinant Human PKC
Cross-reactivity
 <0.5% cross-reactivity observed with available related molecules.<0.5% cross-reactivity observed with available related molecules.
Interference
 No significant interference observed with available related molecules.
 
 

Qualitative Detection
1. Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.
2. Add standard: Set Standard wells,testing sample wells.Add standard 50μl to standard well.
3. Add Sample: Add testing sample 10μl Then add sample diluent 40μl to testing sample well; Blank well doesn’t add anyting.
4. Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°C.
5. Aspirate each well and wash, repeating the process four times for a total of five washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash,remove any remaining Wash Solution by aspirating or decanting. Invert the plate and blot it against clean paper towels.
6. Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.
7. Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.

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