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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.
| Product Name | HSP27 ELISA Kit |
|---|---|
| SKU/Catalog Number | EK7110 |
| Description | Colorimetric detection of HSP27. 96wells/kit, with removable strips. |
| Cite This Product | HSP27 ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7110) |
| Validated Species | Human |
| Application | ELISA *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 Reactivity | There is no detectable cross-reactivity. |
| Pack Size | 96wells/kit, with removable strips. |
| Sensitivity | 0.04 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 Range | 0.2 - 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 Type | Cell 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. |
| Storage | Store at 4°C. |
| Description | Quantity |
| Anti-Hsp27 Immunoassay Plate | 1 Plate |
| 5X Hsp27 Extraction Reagent | 1 vial/10 ml |
| Recombinant Hsp27 Standard | 2 vials |
| Standard and Sample Diluent | 1 vial/ 50 ml |
| 10X Wash Buffer Concentrate | 1 vial/100 ml |
| Anti-Hsp27 Biotinylated Antibody Concentrate | 1 vial/150 ?l |
| Anti-Hsp27 Biotinylated Antibody Diluent | 1 vial/ 13 ml |
| Streptavidin: HRP Concentrate | 1 vial/150 ?l |
| Streptavidin: HRP Diluent | 1 vial/ 13 ml |
| TMB Substrate | 1 vial/ 13 ml |
| Stop Solution | 1 vial/ 13 ml |
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.
You can check the tissue specificity below for information on selecting positive and negative control.
| Gene Name | HSPB1 |
|---|---|
| Protein Name | Heat shock protein beta-1 |
| Protein Function | Small 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 Specificity | Detected 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 Localization | Cytoplasm. |
| Uniprot ID | P04792 |
| Alternative Names | Heat 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 Areas | Cancer, 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.
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.
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.
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.
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