Rat 5-Hydroxytryptamine,5HT ELISA Kit

Code CSB-E08364r
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

This Rat 5HT ELISA Kit was designed for the quantitative measurement of Rat 5HT protein in serum, plasma, cell lysates, tissue homogenates. It is a Competitive ELISA kit, its detection range is 0.8 ng/mL-160 ng/mL and the sensitivity is 0.4 ng/mL.

Target Name 5-Hydroxytryptamine,5HT
Abbreviation 5HT
Species Rattus norvegicus (Rat)
Sample Types serum, plasma, cell lysates, tissue homogenates
Detection Range 0.8 ng/mL-160 ng/mL
Sensitivity 0.4 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Neuroscience
Assay Principle quantitative
Measurement Competitive
Precision
Intra-assay Precision (Precision within an assay): CV%<15%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<15%      
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 rat 5-HT 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)  
1:1 Average % 94  
Range % 89-98  
1:2 Average % 95  
Range % 92-99  
1:4 Average % 97  
Range % 89-103  
1:8 Average % 95  
Range % 90-100  
Recovery
The recovery of rat 5-HT 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) 96 91-102  
EDTA plasma (n=4) 96 92-103  
             
             
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    
160 0.069 0.074 0.072    
40 0.181 0.195 0.188    
12.8 0.426 0.439 0.433    
3.2 0.802 0.855 0.829    
0.8 1.155 1.231 1.193    
0 2.158 2.227 2.193    
Materials provided
  • A 96-well Assay plate -- The 96-well plate has been pre-coated with goat-anti-rabbit 5HT antibody.
  • Standard 6 x 0.5 ml -- Dilute the standard at dilution series, read the OD values, and then draw a standard curve.
  • Anti-rat 5HT Antibody 1 x 6 ml -- Act as the detection antibody.
  • HRP-conjugated 5HT 1 x 6 ml -- Bind to the 5HT antibody, and HRP catalyzes the substrate A and B to elicit a chromogenic reaction.
  • Wash Buffer (20 x concentrate) 1 x 15 ml -- Wash away unbound or free substances.
  • Substrate A 1 x 7 ml -- Mix with substrate B, and the TMB is catalyzed by HRP to develop color.
  • Substrate B 1 x 7 ml -- Mix with substrate A, and the TMB is catalyzed by HRP to develop color.
  • Stop Solution 1 x 7 ml -- Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells)
  • An Instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 600 nm - 630 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
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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