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DNA Damage (8-OHdG) ELISA kit

DNA Damage

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ELISA试剂盒 - Other Elisa Kit

说明书:

EK7008

5083元/96T  

Cell lysates, Plasma, Sample matrices, Urine

ELISA试剂盒(96T)

3-4周

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Product Brief

  • Introduction

    This ELISA kit is of competitive format. Competitive ELISA, also known as inhibition ELISA, is a surface/plate based assay, where the plate is coated with capture antibodies reactive to the molecule of interest. The sample (containing native molecule of interest) and enzyme conjugated recombinant protein (the competing molecule) are added to the coated wells. Since the amount of enzyme conjugated molecule in each well is constant, the level of native molecule in the sample will determine the binding ratio of enzyme conjugated molecule vs. native molecule. After an incubation period, any unbound antibody is washed off. Enzyme substrate (for example, TMB for HRP) is added to each well and will be transformed into a blue precipitate, the amount of which is linearly proportional to the amount of enzyme in the well. The precipitate is then turned into yellow by adding the acid stop solution and the concentration of yellow precipitate is read at 450nm for light absorbance (O.D. value). The O.D. is then used to calculate the amount of molecule of interest in each well, by comparing each sample well against the standard curve. The standard curve is generated using the same principle but instead of adding samples, a series of recombinant molecules with known concentrations are added to 6-8 wells.

    Overview

    Product NameDNA Damage (8-OHdG) ELISA kit
    SKU/Catalog NumberEK7008
    DescriptionCompetitive High Sensitivity ELISA kit for Quantitative Detection of 8-hydroxy-2-deoxy Guanosine. 96wells/kit
    Cite This ProductDNA Damage (8-OHdG) ELISA kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7008)
    Validated SpeciesMouse
    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 the kit at 4°C and -20°C. The reagents are stable until expiration of the kit. Do not expose reagent to heat, sun, or strong light. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)

    Kit Components

    DescriptionQuantity
    8-hydroxy-2-deoxy Guanosine : BSA Coated Plate12x8x1 Microwells
    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. Distilled or deionized water

    2. Precision pipettes

    3. Disposable pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.

    4. ELISA plate reader capable of measuring at 450nm

    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 (1-4). 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(5-9). 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.

Instructions

WARNINGS AND PRECAUTIONS

1. Not for use in humans.

2. Not for use in diagnostics or therapeutics.

3. For in vitro research use only.

4. The components in this kit are intended for use as an integral unit. The components of different lots should not be mixed.

5. It is recommended that standards, control and serum samples be run in duplicate.

6. Optimal results will be obtained by strict adherence to this protocol. Accurate and precise pipetting, as well as following the exact time and temperature requirements prescribed are essential. Any deviation from this may yield invalid data.

ASSAY PROCEDURE

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

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

3. Add 50 uL 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 uL 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 uL 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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