Human advanced glycation end products,AGEs ELISA Kit

Instructions
Code CSB-E09412h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human advanced glycation end products (AGEs) ELISA Kit allows for the in vitro quantitative determination of AGEs concentrations in serum, plasma, or tissue homogenates. This kit exclusively recognizes human AGEs. AGEs are actively produced and accumulated in the circulating blood and various tissues under inflammatory conditions. AGEs also accelerate the expression of receptors for AGEss (RAGEs). The AGEs-RAGEs interaction plays an important role in the development of diabetic vascular complications through the activation of the NF-κB pathway that elicits oxidative stress and inflammation.

The detection mechanism of this kit is based on the Sandwich-ELISA technique. AGEs in the sample is bound to the capture antibody immobilized on the microtiter plate and then sandwiched with Biotin-labeled AGEs antibody. The solution color develops into blue after the ordinal addition of HRP-avidin and TMB. The color development is terminated after adding the stop solution, and the color turns from blue to yellow. The color intensity is positively correlated with AGEs content in the sample. The kit has been quality controlled with high sensitivity, strong specificity, good linearity, precision less than 10%, recovery with 85%-102%, and high lot-to-lot consistency. Refer to the product manual for more information.

Target Name advanced glycation end products,AGEs
Abbreviation AGEs
Species Homo sapiens (Human)
Sample Types serum, plasma, tissue homogenates
Detection Range 0.78 μg/mL-50 μg/mL
Sensitivity 0.195 μg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Immunology
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 human AGEs 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 % 84  
Range % 80-89  
1:2 Average % 90  
Range % 85-95  
1:4 Average % 94  
Range % 90-99  
1:8 Average % 104  
Range % 100-109  
Recovery
The recovery of human AGEs 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) 92 88-96  
EDTA plasma (n=4) 97 92-101  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
µg/ml OD1 OD2 Average Corrected  
50 2.698 2.652 2.675 2.534  
25 2.076 2.082 2.079 1.938  
12.5 1.204 1.283 1.244 1.103  
6.25 0.639 0.669 0.654 0.513  
3.12 0.388 0.374 0.381 0.240  
1.56 0.261 0.259 0.260 0.119  
0.78 0.187 0.179 0.183 0.042  
0 0.145 0.137 0.141    
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with anti-human AGEs antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • 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 AGEs 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 solution.
  • 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 chromogenic reaction.
  • 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 incubation.
  • 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°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

Customer Reviews and Q&A

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 Q&A
Q:

I would like to know if this kit can evaluate all AGEs or only a specific one, like arboximetilisina.

A:
Thanks for your inquiry.
This kit quantifys all AGEs.

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