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Description
Human HSP90b1 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000 × g for 5-10 minutes, and the supernatant can be assayed. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent). Prepare immediately. 5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing). Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return to 4°C. 2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells. Add 100 μL of universal diluent to the blank wells. Cover with a film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.) 3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing. Add 100 μL of Biotinylated Antibody Working Solution directly to each well. Cover with a film and incubate at 37°C for 60 minutes. 4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well. Cover with a film and incubate at 37°C for 30 minutes. 6. Washing: Discard the liquid and wash the plate five times as in step 4. 7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes. 8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at a wavelength of 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest. Multiply the sample concentration by the corresponding dilution factor. |
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| Theory | This kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a capture antibody against Heat Shock Protein 90kDa Beta 1 (HSP90b1). After incubation and washing, the assay is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Heat Shock Protein 90kDa Beta 1 (HSP90b1) in the sample. The absorbance (OD) is measured at 450nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Heat Shock Protein 90kDa Beta 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The 90 kDa heat shock protein β1 (HSP90β1) is a protein encoded by the HSP90B1 gene. This gene encodes a member of the adenosine triphosphate (ATP) metabolic chaperone family, which stabilizes and folds other proteins. The encoded protein is localized to melanosomes and the endoplasmic reticulum. A microRNA gene is located within the 5' exon of this gene. Pseudogenes of this gene are found on chromosomes 1 and 15. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. 2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. 3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. 4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. 5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. 8. Do not use expired products, and do not mix components with different product numbers and batches. 9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. 10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures. |
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| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.4 ★★★★★
Based on 9 reviews
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Product Reviews
★★★★★ 5
Lifesaver Tire Repair Kit Every Car Should Have One
Size: 01- 24pcs (Classic Green)
This 24-piece tire repair kit has already saved me and my family multiple times. My car literally can’t go without it now. It’s compact, well-organized, and the tools are sturdy and professional-grade. Having this kit has saved us from costly trips to the tire shop, time wasted going back and forth, borrowing another car, or risking driving on a flat tire. Seriously, everyone should have one in their car, just buy it and keep it there. Trust me, it’s a game-changer!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 12, 2026
★★★★★ 4
Had no issue and it was very easy to use. It has been a week and no leak.
Size: 01- 24pcs (Classic Green)
Easy to use and its been a week and no leak. The kit is great and easy to use. The hard part was for me to get the screw out of my tire because the head was half gone. Once I got the screw out I followed the instructions and reamed the hole and inserted the plug and trimmed the plug.
If you plan on keeping it in your car you probably should add a screwdriver and/or pliers to get the screw out of the tire and a razor to trim the plug once it is in. You really need a razor to trim the plug I tried with a knife but the had to get a razor.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 5, 2025
★★★★★ 5
Great Item
Size: 01- 24pcs (Classic Green)
Nice kit. Has athe tools and pieces for many easy repairs. Fits neatly inside durable case.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 25, 2026
★★★★★ 5
Quality product
Size: 07- 22pcs (Blue)
Great kit for the price. Reamer and plug tool are strong and durable. Removable handles makes it great for broken tool or putting reamer on a drill if your lazy like me. Easy to use, easy to store and quality plugs
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 30, 2025
★★★★★ 5
Emergency flat tire repair kit
Size: 02- 24pcs (Black)
Experience confidence on every road with the 24 Pcs Professional Heavy Duty Universal Emergency Flat Tire Repair Kit, the ultimate solution for fast, reliable, and long-lasting tire repairs. Designed for cars, motorcycles, trucks, trailers, RVs, ATVs, UTVs, and more, this kit provides everything you need to fix punctures quickly—without calling roadside assistance or waiting for a tow truck.
Built with high-quality, heavy-duty materials, each tool in the set is engineered for durability and performance. The solid steel T-handle reamer and inserter tools offer maximum strength, giving you the leverage and control needed to remove debris and plug punctures with ease. The professional-grade rubber repair plugs are strong, flexible, and designed to create an airtight seal, ensuring your tire holds pressure even in demanding conditions. Whether you’re dealing with a nail, screw, or sharp stone puncture, this kit helps you handle the situation safely and efficiently.
The set includes 24 essential pieces, such as repair plugs, valve stems, valve caps, a pressure gauge, lubricant, and additional accessories that make the repair process smooth and straightforward. Every item fits neatly into the compact, shock-resistant storage case, making it easy to carry in your vehicle’s trunk, glove box, or toolbox. This kit is perfect for emergency situations, long road trips, off-road adventures, work vehicles, and everyday drivers who want to be prepared for anything.
Even if you’re not a mechanic, the repair process is simple. The kit includes all the tools needed to clean, prepare, and seal the puncture, allowing you to get back on the road in minutes. It’s a cost-saving, time-saving, and stress-reducing solution every driver needs.
Stay prepared, stay protected, and stay moving with the 24 Pcs Universal Heavy Duty Tire Repair Kit—a reliable companion designed to handle unexpected flats with professional power and everyday convenience.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 25, 2025
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