Human Hyaluronic acid,HA ELISA Kit

Instructions
Code CSB-E04805h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human Hyaluronic acid (HA) ELISA Kit is designed to measure HA biological samples, including serum, plasma, cerebrospinal fluid (CSF), and tissue homogenates, in a quantitative fashion. This kit has been verified by multiple tests with advantages of high sensitivity, strong specificity, precision less than 10%, and lot-to-lot consistency. Based on the Sandwich-ELISA technique in combination with the enzyme-substrate chromogenic reaction as well as colorimetric measurement, the levels of HA in the sample can be calculated.

HA is a large non-sulfated glycosaminoglycan naturally found in many areas of the human body, including the skin, eyes, and synovial fluid (SF) of the joints. As a humectant, HA retains moisture and plays an important role in the biomechanics of normal SF, where it is partially responsible for lubrication and viscoelasticity of the SF. HA can be used in beauty and skincare products through biofermentation by bacteria.

Target Name Hyaluronic acid,HA
Alternative Names N/A
Abbreviation HA
Species Homo sapiens (Human)
Sample Types serum, plasma, cerebrospinal fluid (CSF), tissue homogenates
Detection Range 0.156 ng/mL-10 ng/mL
Sensitivity 0.039 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Cell Biology
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 HA 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)  
1:20 Average % 87  
Range % 80-94  
1:40 Average % 92  
Range % 88-99  
1:80 Average % 97  
Range % 91-100  
1:160 Average % 101  
Range % 97-105  
Recovery
The recovery of human HA 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) 100 95-105  
EDTA plasma (n=4) 93 88-98  
             
             
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 Corrected  
10 2.727 2.700 2.714 2.626  
5 2.231 2.345 2.288 2.200  
2.5 1.558 1.655 1.607 1.519  
1.25 0.831 0.901 0.866 0.778  
0.625 0.477 0.451 0.464 0.376  
0.312 0.301 0.322 0.312 0.224  
0.156 0.189 0.174 0.182 0.094  
0 0.087 0.089 0.088    
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human HA 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.
  • Biotin-labeled HA antibody(100 x concentrate) 1 x 120 μl ---Act as the detection antibody.
  • HRP-avidin (100 x concentrate) 1 x 120 μl --- Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • Biotin-antibody Diluent 1 x 15 ml ---Dilute the high concentration Biotin-antibody to an appropriate working solution.
  • HRP-avidin Diluent 1 x 15 ml ---Dilute the high concentration HRP-avidin solution to an appropriate solution.
  • Sample Diluent 1 x 50 ml---Dilute the sample to an appropriate concentration.
  • Wash Buffer (25 x concentrate) 1 x 20 ml --- Wash away unbound or free substances.
  • TMB Substrate 1 x 10 ml --- Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • Stop Solution 1 x 10 ml --- 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 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

Customer Reviews and Q&A

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 Q&A
Q:

We are looking into the best method to distinguish HA manufactured by fermentation and nature human HA.

A:
Thanks for your inquiry.
CSB-E04805h Human Hyaluronic acid, HA ELISA Kit
This kit can detect nature HA in polymerized form. As for HA manufactured by fermentation, it usually exists in free form.
We think it can not be detected out by the kit.

  Tel: 301-363-4651
  Email: support@cusabio.com
  Distributors Worldwide

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