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DNA Damage (8-OHdG) ELISA Kit

DNA Damage (8-OHdG)

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ELISA试剂盒 - Other Elisa Kit

说明书:

Typical Standard Curve for the DNA Damage (8-OHdG) ELISA Kit (Enzyme-Linked Immunosorbent Assay)–EK7114 Assay Type: Competitive ELISA. Detection Method: Colorimetric Assay. Assay Range: 0.94 – 60 ng/ml. Chemical Equation of the Oxidation of Guanosine Diagram of the 8-OHdG Competitive ELISA Urine Spike Assay Diagram of the Preparation of the 8-OHdG Standards Diagram of the Triplicate Sample Plate Format Preview of the Calculations Worksheet
  • Typical Standard Curve for the DNA Damage (8-OHdG) ELISA Kit (Enzyme-Linked Immunosorbent Assay)–EK7114 Assay Type: Competitive ELISA. Detection Method: Colorimetric Assay. Assay Range: 0.94 – 60 ng/ml.
  • Chemical Equation of the Oxidation of Guanosine
  • Diagram of the 8-OHdG Competitive ELISA
  • Urine Spike Assay
  • Diagram of the Preparation of the 8-OHdG Standards
  • Diagram of the Triplicate Sample Plate Format
  • Preview of the Calculations Worksheet

EK7114

5070元/96T  

Cell lysates, Plasma, Sample matrices, Urine

ELISA试剂盒(96T)

3-4周

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总价格:

---元*1

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Product Brief

  • Introduction

    Boster's ELISA Kit is a competitive assay that can be used for the quantification of 8-OHdG in urine, cell culture, plasma, and other sample matrices. The ELISA utilizes an 8-hydroxy-2-deoxy Guanosine-coated plate and an HRP-conjugated antibody for detection which allows for an assay range of 0.94 - 60 ng/mL, with a sensitivity of 0.59 ng/mL. The other highlights of this kit are a quick incubation time of 60 minutes, stable reagents, and an easy to use protocol. It is important to note that the 8-OHdG antibody used in this assay recognizes both free 8-OHdG and DNA-incorporated 8-OHdG. Since complex samples such as plasma, cell lysates, and tissues are comprised of mixtures of DNA fragments and free 8-OHdG, concentrations of 8-OHdG reported by ELISA methodology will not coincide with those reported by LC-MS where the single nucleoside is typically measured. This should be kept in mind when analyzing and interpreting experimental results.

    Overview

    Product NameDNA Damage (8-OHdG) ELISA Kit
    SKU/Catalog NumberEK7114
    DescriptionColorimetric detection of DNA Damage (8-OHdG). 96wells/kit, with removable strips.
    Cite This ProductDNA Damage (8-OHdG) ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7114)
    Validated SpeciesAll Animals
    ApplicationELISA

    *Our Boster Guarantee covers the use of this product in the above tested applications.

    Cross Reactivity8-Hydroxy-2-deoxy Guanosine (8-OHdG): 100%. 8-Hydroxy Guanosine (8-OHG): 23%. 8-Hydroxy Guanine (8-oxoG): 23%. Guanosine: <0.01%.
    Pack Size96wells/kit, with removable strips.

    Properties

    Sensitivity0.59 ng/mL
    *Sensitivity, or Lower Limit of Detection (LLD), is the minimum level of target protein the ELISA assay can detect. We measure 20 blank wells and if the O.D. value is 2 standard deviations higher than the blanks' average O.D. the sample can be deemed positive.
    Assay Range0.94 - 60 ng/mL
    *This assay range is determined using common samples. For samples with low target protein concentrations, users can adjust the standard curve to extend the lower limit of assay range.
    Sample TypeCell lysates, Plasma, Sample matrices, Urine

    *The above listed samples are the ones valided with the assay. If you do not see your sample of interest listed, as long as there is enough level of target protein present in the sample, this Picokine? ELISA kit should detect it. 
    **For protocol and tips regarding preparing your sample of interest, please check our ELISA sample preparation guide.
    StorageStore at 4°C and -20°C. (All reagents are stable as supplied at 4°C, except the standard, which should be stored at -20°C. )

    Kit Components

    DescriptionQuantity
    8-hydroxy-2-deoxy Guanosine : BSA Coated Plate1 Plate
    8-hydroxy-2-deoxy Guanosine Standard1 vial/ 100uL
    8-hydroxy-2-deoxy Guanosine HRP Conjugated Monoclonal Antibody1 vial/75uL
    Sample and Standard Diluent1 vial/50mL
    8-hydroxy-2-deoxy Guanosine Antibody Diluent1 vial/13mL
    Wash Buffer Concentrate1 vial/50mL
    TMB Substrate1 vial/ 13mL
    Stop Solution1 vial/ 13mL
    Plate Cover2 covers

    Material Required But Not Provided

    1. A plate reader capable of measuring absorbance at 450 nm.

    2. Adjustable pipettes and a repeat pipettor.

    3. Deionized or distilled water.

    4. Materials used for Sample Preparation.

    Background

    8-hydroxy-2-deoxy Guanosine (8-OH-dG) is produced by the oxidative damage of DNA by reactive oxygen and nitrogen species and serves as an established marker of oxidative stress. Hydroxylation of guanosine occurs in response to both normal metabolic processes and a variety of environmental factors (i.e., anything that increases reactive oxygen and nitrogen species). Increased levels of 8-OH-dG are associated with the aging process as well as with a number of pathological conditions including cancer, diabetes, and hypertension. In complex samples such as plasma, cell lysates, and tissues, 8-OH-dG can exist as either the free nucleoside or incorporated in DNA. Once the blood enters the kidney, free 8-OH-dG is readily filtered into the urine, while larger DNA fragments remain in the bloodstream. Because of the complexity of plasma samples, urine is a more suitable matrix for the measurement of free 8-OH-dG than plasma. Urinary levels of 8-OH-dG range between 2.7-13 ng/mg creatine, while plasma levels of free 8-OH-dG have been reported to be between 4-21 pg/ml as determined by LC-MS.

    DNA Damage (8-OHdG) ELISA Kit Images

    Click the images to enlarge.

    Typical Standard Curve for the DNA Damage (8-OHdG) ELISA Kit (Enzyme-Linked Immunosorbent Assay)–EK7114 Assay Type: Competitive ELISA. Detection Method: Colorimetric Assay. Assay Range: 0.94 – 60 ng/ml.

    Typical Standard Curve for the DNA Damage (8-OHdG) ELISA Kit (Enzyme-Linked Immunosorbent Assay)–EK7114 Assay Type: Competitive ELISA. Detection Method: Colorimetric Assay. Assay Range: 0.94 – 60 ng/ml.

    Chemical Equation of the Oxidation of Guanosine

    Chemical Equation of the Oxidation of Guanosine

    Diagram of the 8-OHdG Competitive ELISA

    Diagram of the 8-OHdG Competitive ELISA

    Urine Spike Assay

    Urine Spike Assay

    Diagram of the Preparation of the 8-OHdG Standards

    Diagram of the Preparation of the 8-OHdG Standards

    Diagram of the Triplicate Sample Plate Format

    Diagram of the Triplicate Sample Plate Format

    Preview of the Calculations Worksheet

    Preview of the Calculations Worksheet

Instructions

Intra/Inter Assay Precision

Intra-Assay Precision: Three samples of known concentration were assayed thirty times on one plate; the intra-assay coefficient of variation of the DNA Damage ELISA has been determined to be <5%.
Inter-Assay Precision: Three samples of known concentration were assayed thirty times in three individual assays; the inter-assay coefficient of variation of the DNA Damage ELISA has been determined to be <5%.

Assay Overview

1. Prepare standard and samples in the Sample and Standard Diluent.

2. Add 50 ?L of prepared standards and samples in triplicate to appropriate wells.

3. Add 50 ?L of the diluted antibody preparation to the appropriate wells.


4. Cover plate with Plate Cover and incubate at room temperature (20-25°C) for 1 hour.

5. Wash plate 4 times with 1X Wash Buffer.

6. Add 100 ?L of TMB Substrate to each well.

7. Cover plate and develop the plate in the dark at room temperature for 30 minutes.

8. Add 100 ?L of Stop Solution to each well. <

9. Measure absorbance on a plate reader at 450 nm.

10. Plot the standard curve and calculate sample concentrations.


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