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Description
Human HSF1 ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 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 Transcription Factor 1 (HSF1). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Heat Shock Transcription Factor 1 (HSF1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Heat Shock Transcription Factor 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Heat shock transcription factor 1 (HSF1) is a protein encoded by the HSF1 gene. Highly conserved across eukaryotes, HSF1 is a major mediator of transcriptional responses to proteotoxic stress and plays important roles in non-stress regulation, such as development and metabolism. Diseases associated with HSF1 include heat shock RNA 1 and prion disease. Pathways associated with HSF1 include cellular responses to heat stress and ERK signaling. Gene Ontology (GO) annotations associated with this gene include DNA-bound transcription factor activity and chromatin binding. An important homologue of this gene is HSF4. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.0 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 3
More soft than firm
Size: Queen(Pack of 2)
Not firm but more soft, would I be returning probably not but if you were already on the fence this is your confirmation that it’s not as described, ( well to my standards at least ) it does feel of good quality but I was looking more a sturdy firm that soft and cushion feeling .
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Reviewed in the United States on January 1, 2026
★★★★★ 5
Best fluffy but firm pillows
Size: King(Pack of 2), Size: King(Pack of 2)
These pillows are worth every penny! They started off pretty flat but after three days they plumped up well. In addition to them being plump, they are also firm. Exactly what I was looking for. You can shake up the internal stuffing if you want them softer too. They were a bit pricey but I’m going to repurchase and replace all my home pillows with these. I do sleep hot and the cooling side is perfect to lay on. Kept me cool and comfortable. I haven’t had any neck or back pain after switching to these pillows too. Absolutely recommend these for your home. The photo shows how full the pillows are. I have four pillows in the picture plus two smaller other brands. These are definitely fluffier.
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Reviewed in the United States on January 8, 2026
★★★★★ 5
This Pillow Is A Must Buy!! What Are You Waiting For?
Size: Queen (Pack of 1)
This is, without a doubt, the best pillow I have ever owned. One of my favorite features is the adjustable shredded memory foam fill, which allows you to customize the loft and firmness by adding or removing foam to suit your sleeping preferences. As both a side and back sleeper, I found it incredibly comfortable and supportive.
The overall quality is excellent, from the materials used in the pillow itself to the thoughtful packaging. The removable cover is washable, although I personally use a pillowcase to keep it protected. I also appreciated that extra foam is included, making it easy to fine-tune the pillow over time.
The difference in my sleep quality was noticeable from the very first night. I’ve been sleeping more comfortably and waking up feeling more rested ever since I started using it. Considering the quality, comfort, and customization options, I believe the $75 price point is well worth it. I am so impressed with this pillow that I plan to purchase another one in the future.
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Reviewed in the United States on June 4, 2026
★★★★★ 5
This is a high-quality, well-designed pillow that combines softness, comfort, and adjustability
Size: Queen (Pack of 1), Size: Queen (Pack of 1)
I purchased the DreamyBlue Signature Adjustable Pillow for my husband because he often experiences back discomfort, and many of the pillows we had previously would become flat and lose their support over time.
This pillow has exceeded our expectations. It is incredibly soft, comfortable, and comes with extra filling, which allows you to adjust the firmness to your personal preference. We really appreciated having that option.
Even after extended use, the pillow has maintained its shape much better than other pillows we've owned. It provides a plush, cozy feel while still offering good support. My husband finds it very comfortable, and it has quickly become his favorite pillow.
This is a high-quality, well-designed pillow that combines softness, comfort, and adjustability. I would definitely recommend it.
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Reviewed in the United States on June 4, 2026
★★★★★ 5
Finally Found a Pillow Worth Keeping
Size: Queen (Pack of 1)
I genuinely feel like I’ve tried every pillow on the market. At one point, I thought Amazon was going to flag my account because I bought and returned so many pillows trying to find the right one.
This is the only pillow that made the cut.
The shredded memory foam filling makes a huge difference because the pillow is both soft and supportive, while still being adjustable to fit the way you sleep. It arrives extremely full and plush, and it even comes with an extra bag of fill, which honestly surprised me. I ended up removing about half of the filling to get it exactly how I wanted it.
I’m a side/stomach sleeper, which can make finding a good pillow difficult since most are either too thick or too flat. This one was perfect because I could shape it exactly how I sleep.
It stays cool throughout the night, has a slightly heavier premium feel, and feels much higher quality than most pillows I’ve tried. Super comfortable, supportive, and customizable.
I would absolutely recommend it and would definitely buy it again
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Reviewed in the United States on May 24, 2026
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