8-OHdG(8-Hydroxydeoxyguanosine) ELISA Kit

Category:

This kit is based on Competitive-ELISA detection method and takes 2h assay time. The microplate provided in this kit has been precoated with 8-OHdG. 8-OHdG in the sample or standard competes with a fixed amount of biotin-labeled 8-OHdG for sites on a pre-coated antibody specific to 8-OHdG. Free components are washed off. After adding HRP-Streptavidin Conjugate (SABC), biotin specifically binds with streptavidin to form immune complexes. Wash off unbound conjugate. Add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Compare OD450 value of sample with the standard curve through curve fitting software to determine the concentration of 8-OHdG in the sample. The concentration of the target substance is inversely proportional to the OD450 value.

Catalogue No.: 
EU2548-HS
Alias: 
8-OHdG, 8-Hydroxydeoxyguanosine ELISA Kit
Species: 
Universal
Range: 
0.468-30ng/ml
Sensitivity: 
0.281ng/ml
  • SPECIFICATIONS
Product Name
8-OHdG(8-Hydroxydeoxyguanosine) ELISA Kit
Alias
8-OHdG, 8-Hydroxydeoxyguanosine ELISA Kit
Catalogue No.
EU2548-HS
Size
48T/96T
Species
Universal
Sample Type
Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Urine, Other liquid samples
Detection Method
Competitive ELISA, Coated with Antigen
Detection Wavelength
OD450
Reaction Duration
2 hours
Range
0.468-30ng/ml
Sensitivity
0.281ng/ml
Storage
2-8°C(Sealed), Don't cryopreserve.
Specificity
Specifically binds with 8-OHdG , no obvious cross reaction with other analogues.
ELISA Kit Components
Kit Components Item Size(48T) Size(96T) Storage Condition for Opened Kit
E001 ELISA Microplate(Dismountable) 8×6 8×12 Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E002 Lyophilized Standard 1vial 2vial Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E003 Lyophilized Biotin-labeled Antibody(Concentrated) 1vial 1vial
E034 HRP-Streptavidin Conjugate(SABC, 100X) 60ul 120ul 2-8°C (Avoid Direct Light)
E024 TMB Substrate 5ml 10ml
E005 Purified Water 200ul 200ul 2-8°C
E039 Sample Dilution Buffer 10ml 20ml
E040 Antibody Dilution Buffer 5ml 10ml
E049 SABC Dilution Buffer 5ml 10ml
E026 Stop Solution 5ml 10ml
E038 Wash Buffer(Concentrated, 25X) 15ml 30ml
E006 Plate Sealer 3 pieces 5 pieces  
E007 Product Description 1 copy 1 copy
Required Instruments and Reagents
  1. Microplate reader (wavelength: 450nm)
  2. 37°C incubator (CO2 incubator for cell culture is not recommenced.)
  3. Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
  4. Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
  5. Sterile tubes and Eppendorf tubes with disposable tips
  6. Absorbent paper and loading slot
  7. Deionized or distilled water
Assay Procedure Summary
Elisa实验原理图
  • Step 1: Wash the plate twice before adding the standard and sample.
  • Step 2: Add 50ul standard or sample into each well. Immediately add 50ul biotin-labeled antibody working solution into each well(The tip can't be reused after touching the liquid.), gently shake the plate for 1min, seal closely and statically incubate for 45 minutes at 37°C.
  • Washing: Wash the plate three times and immerse for 1min each time.
  • Step 3: Add 100ul HRP-Streptavidin Conjugate (SABC) working solution into each well, seal the plate and statically incubate for 30 minutes at 37°C.
  • Washing: Wash the plate five times and immerse for 1min each time.
  • Step 4: Add 90ul TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
  • Step 5: Add 50ul stop solution. Read at 450nm immediately and calculate.
Standard Curve

This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.)

Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.

STD.(ng/ml) OD-1 OD-2 Average
0 2.169 2.231 2.2
0.469 1.881 1.935 1.908
0.938 1.484 1.528 1.506
1.875 1.337 1.375 1.356
3.75 0.999 1.029 1.014
7.5 0.612 0.63 0.621
15 0.332 0.342 0.337
30 0.158 0.162 0.16
EU2548-HS Standard Curve Image
Recovery

Add a certain amount of 8-OHdG into the sample. Calculate the recovery by comparing the measured value with the expected amount of 8-OHdG in the sample.

Sample Type Recovery Range(%) Average(%)
serum(n=10) 87-99 92
EDTA plasma(n=10) 92-100 98
Heparin plasma(n=10) 86-104 92
Linearity

Dilute the sample with a certain amount of 8-OHdG at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type 1:2 1:4 1:8
serum(n=10) 85-104% 82-95% 82-97%
EDTA plasma(n=10) 94-102% 92-101% 85-96%
Heparin plasma(n=10) 87-104% 82-101% 85-99%
Precision(%)

Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.

Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.

Item Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/ml) 0.88 3.67 14.75 0.95 3.77 14.34
Standard deviation 0.04 0.18 0.63 0.05 0.2 0.68
CV(%) 5.03 4.84 4.24 5.36 5.2 4.71
Stability

Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.

ELISA kit(n=5) 37°C for 1 month 2-8°C for 6 months
Average(%) 80 95-100