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HSP70 High Sensitivity ELISA kit

HSP70 High Sensitivity

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ELISA试剂盒 - Other Elisa Kit

说明书:

EK7009

7969元/96T  

Cell lysates, Plasma, Serum, Tissue

ELISA试剂盒(96T)

3-4周

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Product Brief

  • Introduction

    This ELISA kit is of sandwich format. A sandwich ELISA measures antigen between two layers of antibodies (capture and detection antibody). 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 NameHSP70 High Sensitivity ELISA kit
    SKU/Catalog NumberEK7009
    DescriptionSandwich High Sensitivity ELISA kit for Quantitative Detection of HSP70. 96wells/kit
    Cite This ProductHSP70 High Sensitivity ELISA kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7009)
    Validated SpeciesMouse
    ApplicationELISA

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

    Cross ReactivityThere is no detectable cross-reactivity.
    Pack Size96wells/kit, with removable strips.

    Properties

    Sensitivity0.02 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 Range0.55 - 35 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, Serum, Tissue

    *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. 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
    Anti-Hsp70 Immunoassay Plate12x8x1 Microwells
    Recombinant Hsp70 Standard2 vials
    Standard and Sample Diluent1 vial/ 50 ml
    10X Wash Buffer Concentrate1 vial/100 ml
    Anti-Hsp70 Biotinylated Antibody Concentrate1 vial/150 ?l
    Anti-Hsp70 Biotinylated Antibody Diluent1 vial/ 13 ml
    Streptavidin: HRP Concentrate1 vial/50 ?l
    Streptavidin: HRP Diluent1 vial/ 13 ml
    TMB Substrate1 vial/ 13 ml
    Stop Solution1 vial/ 13 ml

    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

    HSP70 genes encode abundant heat-inducible 70-kDa HSPs (HSP70s). In most eukaryotes HSP70 genes exist as part of a multigene family. They are found in most cellular compartments of eukaryotes including nuclei, mitochondria, chloroplasts, the endoplasmic reticulum and the cytosol, as well as in bacteria. The genes show a high degree of conservation, having at least 5O% identity. The N-terminal two thirds of HSP70s are more conserved than the C-terminal third. HSP70 binds ATP with high affinity and possesses a weak ATPase activity which can be stimulated by binding to unfolded proteins and synthetic peptides. When HSC70 (constitutively expressed) present in mammalian cells was truncated, ATP binding activity was found to reside in an N-terminal fragment of 44kDa which lacked peptide binding capacity. Polypeptide binding ability therefore resided within the C-terminal half. The structure of this ATP binding domain displays multiple features of nucleotide binding proteins. All HSP70s, regardless of location, bind proteins, particularly unfolded ones. The molecular chaperones of the HSP70 family recognize and bind to nascent polypeptide chains as well as partially folded intermediates of proteins preventing their aggregation and misfolding. The binding of ATP triggers a critical conformational change leading to the release of the bound substrate protein. The universal ability of HSP70s to undergo cycles of binding to and release from hydrophobic stretches of partially unfolded proteins determines their role in a great variety of vital intracellular functions such as protein synthesis, protein folding and oligomerization and protein transport.

    HSP70 High Sensitivity 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 Standard and Sample Diluent.

2. Add 100 uL of Standard or sample to appropriate wells.

3. Cover plate with Plate Sealer and incubate at 37°C for 2 hours.

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

5. Add 100 uL of Biotinylated Antibody Working Solution to each well.

6. Cover plate with Plate Sealer and incubate at at 37°C for 2 hours.

7. Wash plate four times with 1X Wash Buffer.

8. Add 100 uL of Streptavidin-HRP Working Solution to each well.

9. Cover plate with Plate Sealer and incubate at 37°C for 30 minutes.

10. Wash plate four times with 1X Wash Buffer.

11. Add 100 uL of TMB Substrate to each well.

12. Develop the plate in the dark at room temperature for 30 minutes.

13. Stop reaction by adding 100 uL of Stop Solution to each well.

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


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