Rat Testosterone,T ELISA Kit

Instructions
Code CSB-E05100r
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Rat Testosterone/T ELISA Kit is designed for the quantitative determination of endogenic rat testosterone concentrations in serum, plasma, cell culture supernates, or tissue homogenates. This assay employs the competitive inhibition enzyme immunoassay technique. Standards or samples are added to the appropriate goat-anti-rabbit testosterone antibody-pre-coated microtiter plate wells with the anti-rat testosterone antibody and HRP-conjugate. The competitive inhibition reaction is launched between with HRP labeled testosterone and unlabeled testosterone in the sample or standard with the anti-rat Testosterone antibody. Substrate A and B solution are mixed and then added to the wells, and the color turns blue. The color development is stopped, and the intensity of the color is opposite to the amount of DHT in the sample. Read the OD value with the Microplate reader and calculated the DHT concentration referring to the standard curve. This kit has been quality tested by multiple criteria, including sensitivity, specificity, precision, linearity, recovery, and lot-to-lot consistency. Click the product instructions for more information.

Testosterone is a hormone produced by the testes and the adrenal gland. It plays an important role in muscle mass, bone strength, hair growth, and sexual function. Low levels of testosterone can cause low energy, poor concentration, depression, low libido, erectile dysfunction, and other symptoms.

Target Name Testosterone,T
Alternative Names N/A
Abbreviation T
Species Rattus norvegicus (Rat)
Sample Types serum, plasma, cell culture supernates, tissue homogenates
Detection Range 0.13 ng/mL-25.6 ng/mL
Sensitivity 0.06 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Signal Transduction
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 testosteronen 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 % 91  
Range % 86-96  
1:2 Average % 97  
Range % 92-104  
1:4 Average % 85  
Range % 81-88  
1:8 Average % 89  
Range % 83-93  
Recovery
The recovery of rat testosterone 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 87-95  
EDTA plasma (n=4) 94 90-98  
             
             
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    
25.6 0.238 0.236 0.237    
6.4 0.435 0.431 0.433    
2 0.611 0.615 0.613    
0.5 0.984 0.982 0.983    
0.13 1.403 1.398 1.401    
0 2.001 2.003 2.002    
Materials provided
  • A 96-well Assay plate -- The 96-well plate has been pre-coated with goat-anti-rabbit Testosterone antibody.
  • Standard 6 x 0.5 ml -- Dilute the standard at dilution series, read the OD values, and then draw a standard curve.
  • Anti-Testosterone Antibody 1 x 6 ml -- Act as the detection antibody.
  • HRP-conjugated Testosterone 1 x 6 ml -- Bind to the DHT 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
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

Citations

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