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HSP27 ELISA Kit

HSPB1

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ELISA试剂盒 - Other Elisa Kit

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Typical Standard Curve for the HSP27 ELISA Kit (Enzyme-Linked Immunosorbent Assay)–EK7110 Assay Type: Sandwich ELISA. Detection Method: Colorimetric Assay. Assay Range: 0.2 – 13 ng/mL.
  • Typical Standard Curve for the HSP27 ELISA Kit (Enzyme-Linked Immunosorbent Assay)–EK7110 Assay Type: Sandwich ELISA. Detection Method: Colorimetric Assay. Assay Range: 0.2 – 13 ng/mL.

EK7110

6630元/96T  

Cell lysates, Plasma, Serum, Tissue

ELISA试剂盒(96T)

3-4周

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Product Brief

  • Introduction

    Boster's ELISA Kit is for the detection of free, unbound human Hsp27 in cell lysates, tissue extracts, and human serum samples. The latter matrix has significant quantities of Hsp27-complexing components which can obscure the detection of Hsp27 by this assay. Each kit contains sufficient components to quantitate the Hsp27 concentration in up to 40 samples, tested in duplicate.

    Overview

    Product NameHSP27 ELISA Kit
    SKU/Catalog NumberEK7110
    DescriptionColorimetric detection of HSP27. 96wells/kit, with removable strips.
    Cite This ProductHSP27 ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7110)
    Validated SpeciesHuman
    ApplicationELISA

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

    **For positive and negative control design, consult "Tissue specificity" under Protein Target Info.

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

    Properties

    Sensitivity0.04 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 Range0.2 - 13 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 at 4°C.

    Kit Components

    DescriptionQuantity
    Anti-Hsp27 Immunoassay Plate1 Plate
    5X Hsp27 Extraction Reagent1 vial/10 ml
    Recombinant Hsp27 Standard2 vials
    Standard and Sample Diluent1 vial/ 50 ml
    10X Wash Buffer Concentrate1 vial/100 ml
    Anti-Hsp27 Biotinylated Antibody Concentrate1 vial/150 ?l
    Anti-Hsp27 Biotinylated Antibody Diluent1 vial/ 13 ml
    Streptavidin: HRP Concentrate1 vial/150 ?l
    Streptavidin: HRP Diluent1 vial/ 13 ml
    TMB Substrate1 vial/ 13 ml
    Stop Solution1 vial/ 13 ml

    Material Required But Not Provided

    1. Ultra pure water.

    2. Additional reagents and materials for cell lysate and tissue extract preparation, including protease inhibitors.

    3. Precision pipettors, with disposable plastic tips.

    4. Polypropylene or polyethylene tubes to prepare samples ? do not use polystyrene, polycarbonate or glass tubes.

    5. A container to prepare 1X Wash Buffer.

    6. A wash bottle or an automated 96-well plate washer.

    7. Disposable reagent reservoirs.

    8. A standard microtiter plate reader for measuring absorbance at 450 nm.

    9. Adhesive plate sealers.

    Protein Target Info (Source: Uniprot.org)

    You can check the tissue specificity below for information on selecting positive and negative control.

    Gene NameHSPB1
    Protein NameHeat shock protein beta-1
    Protein FunctionSmall heat shock protein which functions as a molecular chaperone probably maintaining denatured proteins in a folding- competent state (PubMed:10383393, PubMed:20178975). Plays a role in stress resistance and actin organization (PubMed:19166925). Through its molecular chaperone activity may regulate numerous biological processes including the phosphorylation and the axonal transport of neurofilament proteins (PubMed:23728742).
    Tissue SpecificityDetected in all tissues tested: skeletal muscle, heart, aorta, large intestine, small intestine, stomach, esophagus, bladder, adrenal gland, thyroid, pancreas, testis, adipose tissue, kidney, liver, spleen, cerebral cortex, blood serum and cerebrospinal fluid. Highest levels are found in the heart and in tissues composed of striated and smooth muscle.
    Subcellular LocalizationCytoplasm.
    Uniprot IDP04792
    Alternative NamesHeat shock protein beta-1; HspB1; 28 kDa heat shock protein; Estrogen-regulated 24 kDa protein; Heat shock 27 kDa protein; HSP 27; Stress-responsive protein 27; SRP27; HSPB1; HSP27; HSP28
    Research AreasCancer, Actin, Actin Assembly, Atherosclerosis, Cardiovascular System, Cell Signaling, Chaperone Proteins, Contractility, Cytoskeleton, Heart, Microfilaments, Microtubules, Protein Trafficking, Tumor Biomarkers|

    *if product is indicated to react with multiple species, protein info is based on the human gene.

    Background for Heat shock protein beta-1

    HSP27s belong to an abundant and ubiquitous family of small heat shock proteins (sHSP). It is an important HSP found in both normal human cells and cancer cells. The basic structure of most sHSPs is a homologous and highly conserved amino acid sequence, with an alpha-crystallin-domain at the C-terminus and the WD/EPF domain at the less conserved N-terminus. This N-terminus is essential for the development of high molecular oligomers. HSP27-oligomers consist of stable dimers formed by as many as 8-40 HSP27 protein monomers. The oligomerization status is connected with the chaperone activity: aggregates of large oligomers have high chaperone activity, whereas dimers have no chaperone activity. HSP27 is localized to the cytoplasm of unstressed cells but can redistribute to the nucleus in response to stress, where it may function to stabilize DNA and/or the nuclear membrane. Other functions include chaperone activity (as mentioned above), thermotolerance invivo, inhibition of apoptosis, and signal transduction. Specifically, in vitro, it acts as an ATP-independent chaperone by inhibiting protein aggregation and by stabilizing partially denatured proteins, which ensures refolding of the HSP70 complex. HSP27 is also involved in the apoptotic signaling pathway because it interferes with the activation of cytochrome c/Apaf-1/dATP complex, thereby inhibiting the activation of procaspase-9. It is also hypothesized that HSP27 may serve some role in cross-bridge formation between actin and myosin. And finally, HSP27 is also thought to be involved in the process of cell differentiation. The up-regulation of HSP27 correlates with the rate of phosphorylation and with an increase of large oligomers. It is possible that HSP27 may play a crucial role in termination of growth.

    HSP27 ELISA Kit Images

    Click the images to enlarge.

    Typical Standard Curve for the HSP27 ELISA Kit (Enzyme-Linked Immunosorbent Assay)–EK7110 Assay Type: Sandwich ELISA. Detection Method: Colorimetric Assay. Assay Range: 0.2 – 13 ng/mL.

    Typical Standard Curve for the HSP27 ELISA Kit (Enzyme-Linked Immunosorbent Assay)–EK7110 Assay Type: Sandwich ELISA. Detection Method: Colorimetric Assay. Assay Range: 0.2 – 13 ng/mL.

Instructions

Assay Overview

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

2. Add 100 ?L of Standard or sample to appropriate wells.

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

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

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

6. Cover plate with Plate Sealer and incubate at room temperature for 1 hour.

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

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

9. Cover plate with Plate Sealer and incubate at room temperature for 30 minutes.

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

11. Add 100 ?L 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 ?L of Stop Solution to each well.

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


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