Human Glycated hemoglobin A1c,GHbA1c ELISA Kit

Instructions
Code CSB-E08139h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human Glycated hemoglobin A1c (GHbA1c) ELISA Kit is suitable for the quantitative measurement of GHbA1c in lysates for RBC (red blood cell). This kit is easy to operate and can help to streamline the experimental procedures. It has undergone rigorous quality control in multiple parameters, including specificity, sensitivity, precision, linearity, and lot-to-lot consistency.

The detection mechanism is based on the direct competitive inhibition enzyme immunoassay technique. The microplate has been pre-coated antibody specific for GHbA1c. Standards and samples are pipetted into the wells with Biotin-conjugated GHbA1c. A competitive inhibition reaction is launched between GHbA1c (Standards or samples) and Biotin-conjugated GHbA1c with the pre-coated GHbA1c antibody. After washing, HRP-avidin is added to the wells. Following a wash to remove any unbound reagent, the substrate solution is added to the wells and color develops in opposite to the amount of GHbA1c bound in the initial step. The color development is stopped and the intensity of the color is measured.

GHbA1c is a sugar-bound hemoglobin that is formed through the non-enzymatic binding of circulating glucose to hemoglobin. This process is known as glycation. The rate of GHbA1c formation is positively correlated with the surrounding blood glucose concentration. GHbA1c is a remarkable indicator for Diabetes Mellitus (DM) patients and can evaluate long-term average glycemia.

Target Name Glycated hemoglobin A1c,GHbA1c
Abbreviation GHbA1c
Species Homo sapiens (Human)
Sample Types lysates for RBC
Detection Range 0.012 ng/mL-50 ng/mL
Sensitivity 0.012 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Others
Assay Principle quantitative
Measurement Competitive
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.    
             
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    
50 0.270 0.273 0.272    
12.5 0.367 0.378 0.373    
3.125 0.463 0.489 0.476    
0.78 0.560 0.598 0.579    
0.195 0.692 0.672 0.682    
0.049 1.384 1.368 1.376    
0.012 1.994 1.954 1.974    
0 2.373 2.399 2.386    
Materials provided
  • A 96-well Assay plate --The 96-well plate has been pre-coated with GHbA1c antibody.
  • Standard (10 x concentrate) (1 x 200 μl) --Dilute the standard at dilution series, read the OD values, and then draw a standard curve.
  • Biotin-conjugated GHbA1c (100 x concentrate) (1 x 60 μl)
  • HRP-avidin (100 x concentrate) (1 x 120 μl) --Catalyze the TMB solution, eliciting a chromogemic reaction.
  • Biotin-conjugate Diluent (1 x 10 ml) --Dilute the Biotin-conjugated GHbA1c
  • HRP-avidin Diluent (1 x 20 ml) --Dilute the HRP-avidin
  • Sample Diluent (2 x 20 ml) --Reconstitute the standard and dilute the sample to an appropriate concentration.
  • Wash Buffer (25x concentrate) (1 x 20 ml)--Wash away unbound or free substances.
  • TMB Substrate (1x 10 ml) --Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • Stop Solution (1 x 10ml) --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 540 nm - 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

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