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Boster's ELISA Kit is for the detection of human Hsp90α in cell lysates, tissue extracts, and serum samples. Each kit contains sufficient components to quantitate the Hsp90α concentration in up to 40 samples, tested in duplicate. The ELISA is specific for Hsp90α and does not cross react with Hsp90α, Grp94, Hsp60 or Hsp70.
| Product Name | HSP90 alpha ELISA Kit |
|---|---|
| SKU/Catalog Number | EK7108 |
| Description | Colorimetric detection of HSP90 alpha. 96wells/kit, with removable strips. |
| Cite This Product | HSP90 alpha ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7108) |
| Validated Species | Goat, Human |
| Application | ELISA *Our Boster Guarantee covers the use of this product in the above tested applications. |
| Cross Reactivity | There is no detectable cross-reactivity. |
| Pack Size | 96wells/kit, with removable strips. |
| Sensitivity | 0.117 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 | 0.44 - 28 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, 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-Hsp90a Immunoassay Plate | 1 Plate |
| 5X Hsp90a Extraction Reagent | 1 vial/10 ml |
| Recombinant Hsp90a Standard | 2 vials |
| Standard and Sample Diluent | 1 vial/ 50 ml |
| 10X Wash Buffer Concentrate | 1 vial/100 ml |
| Anti-Hsp90a Biotinylated Antibody Concentrate | 1 vial/150 ?l |
| Anti-Hsp90a 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.
HSP90 is a highly conserved and essential stress protein that is expressed in all eukaryotic cells. From a functional perspective, HSP90 participates in the folding, assembly, maturation, and stabilization of specific proteins as an integral component of a chaperone complex. Despite its label of being a heat-shock protein, HSP90 is one of the most highly expressed proteins in unstressed cells (1–2% of cytosolic protein). It carries out a number of housekeeping functions – including controlling the activity, turnover, and trafficking of a variety of proteins. Most of the HSP90- regulated proteins that have been discovered to date are involved in cell signaling. The number of proteins now known to interact with HSP90 is about 100. Target proteins include the kinases v-Src, Wee1, and c-Raf, transcriptional regulators such as p53 and steroid receptors, and the polymerases of the hepatitis B virus and telomerase. When bound to ATP, HSP90 interacts with co-chaperones Cdc37, p23, and an assortment of immunophilin-like proteins, forming a complex that stabilizes and protects target proteins from proteasomal degradation. In most cases, HSP90-interacting proteins have been shown to co-precipitate with HSP90 when carrying out immune-oadsorption studies, and to exist in cytosolic heterocomplexes with it. In a number of cases, variations in HSP90 expression or HSP90 mutation has been shown to degrade signaling function via the protein or to impair a specific function of the protein (such as steroid binding, kinase activity) in vivo. Ansamycin antibiotics, such as geldanamycin and radicicol, inhibit HSP90 function.
Click the images to enlarge.
Typical Standard Curve for the HSP90 Alpha ELISA kit (Enzyme-Linked Immunosorbent Assay) – EK7108 Assay Type: Sandwich ELISA. Detection Method: Colorimetric Assay. Assay Range: 0.44 – 28 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 37°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, 20-25 °C 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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