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Description
Human HPA 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: 1. 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: 1. 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. 2. 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. 3. 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. 4. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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 heparanase (HPA) capture antibodies. After incubation and washing, the sample 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 heparanase (HPA) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Heparanase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Heparanase (HPA), also known as HPSE, is an enzyme that acts on the cell surface and within the extracellular matrix, degrading polymerized heparan sulfate molecules into shorter oligosaccharides. This protein is initially synthesized in the Golgi apparatus as an inactive 65 kDa proheparinase and translocated to late endosomes/lysosomes for transport to the cell surface. Within lysosomes, it is processed into its active form by proteases. It has endoglycosidase activity, cleaving polymerized heparan sulfate molecules at sites within the polymer chain. This enzyme degrades the heparan sulfate scaffolding of the basement membrane and extracellular matrix. It is also implicated in inflammatory processes, allowing extravasation of activated T lymphocytes. In ocular surface physiology, this activity serves as a switch for prosecretory mitogens. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.1 ★★★★★
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Product Reviews
★★★★★ 1
Blades are not sharp and will not remove peach fuzz, even after two swipes
Color: #10 - Pack of 21 (20 Blades + 1 Handle)
I was really disappointed in this. It did not remove all of the peach fuzz completely and I dermaplaned twice back to back. Which tells me the blades aren’t as sharp as they’re made out to be. The blade handle was sturdy and seemed to be made of good quality. But the blades themselves are horrible at cutting and aren’t razor sharp. I personally wouldn’t purchase this if I were you. I’ll keep the blade handle, but I will definitely be purchasing different blades.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 14, 2026
★★★★★ 5
Good quality!
Color: #14R - Pack of 21 (20 Blades + 1 Handle)
If you are looking to dermaplane at home obviously be careful but if you are looking into getting scalpels, these are good quality and come in a big pack where it will last you a while! Remember dermaplaning is once every four weeks and make sure you have no active breakouts!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 14, 2025
★★★★★ 3
Not the sharpest blades
Not fantastic but they work
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 15, 2026
★★★★★ 5
The BEST relationship book you will ever read! Single, Dating, or Married this book will transform your relationships forever!
Format: Paperback
This book is incredible! Debra's insights as a counselor are unlike anything I have ever read and will change my relationships forever. The survey results from 1000 singles/1000 couples were really eye opening, but more importantly she presented solutions and offered hope in many areas: dealing with unmet expectations, how to be authentic, how to protect your marriage, dealing with conflict, issues with sex, how to truly become "one", and many more.
Many relationship books are about a couples' journey or their advice that worked for them, Debra is unique in that your are getting perspective from a licensed counselor and relationship expert who has worked with countless clients! Her wisdom shines through and her approach to a number of subjects are so unique, refreshing, and inspiring. She does a masterful job of approaching relationships with her clinical wisdom, but rooted in her deep love for Jesus. I wish someone would have told or taught me countless things in this book years ago.
I loved reading her stories of couples and singles she worked with, many real-life examples made the book very connectable and authentic. My favorite chapters were Alter that Ego, From Pride to Humility (where you actually learn what a proper apology should look like in a relationship and the power of humility in a relationship); Sex Marks the Spot, From Infatuation to Adoration (amazing chapter about navigating sex and dealing with problem spots); and Walls Will Fall, From Vulnerability to Intimacy (
If you are single, READ this book to get a front row seat into what marriage will look like and more importantly the qualities you should be looking for in your future spouse!
if you are dating/engaged READ this book to learn more about your relationship, future areas and topics you need to discuss and more importantly how to handle them.
if you are married (whether for a year or 50 yrs) READ this book to transform your marriage, Debra will equip you with the tools to deal with issues you may be struggling with and even more how to identity them.
I walked away from this book so encouraged and filled with hope as I got a glimpse of what God's best in relationship should look like.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 1, 2018
★★★★★ 5
Choosing Marriage
Format: Paperback
I don't remember when I first discovered Debra Fileta, but she is my soul sister. Debra is a self-proclaimed realist, which is why I like her approach so much. She is the author of the site, True Love Dates. She does not beat around the bush with her relationship advice.
I signed up to be on the book's Launch team, but I was quickly reminded I should not plan to do anything at the end of the school year - oops - my bad! So sorry, Debra. Better late than never, right?!
I am a slow reader, but I wanted to make sure I was taking the time to gain the wisdom and knowledge Debra was sharing with singles and couples alike about relationships. Each chapter ended with reflection questions for singles and couples.
Now to get to why you all should grab a copy from Amazon or your local bookstore - it is AWESOME. Worth it. I like to read relationship books because I am far from perfect and like to hear what others have to say on the subject.
"A good marriage comes from a series of decisions over an extended period of time, with the commitment of two people choosing to love each other no matter what my come their way" (23). I wish people could understand this concept - and I am saying this as a single person. Even though I have seen the good, the bad, and the ugly of marriages around me, it still remains a life goal to be a wife and mother. I believe it is worth it and reading this book has given me hope that it is possible to have a good marriage. I will never truly understand marriage until I am married because "[y]ou can't experience real love with a person until you've experienced the entirety of a person - the good, the bad, the ugly, and the beautiful" (101).
In this book, Debra addresses communication barriers, boundaries, and learning about good sex within marriage. She shares personal stories as well as stories from those she counseled.
Debra is a straight-shooter when she states, "Seek accountability.... Get a tight rein on your thought life! Stop feeding your mind entertainment that will pollute it with lust.... [I]f you don't have a community, it's time to turn off Netflix, get off social media, and start investing in real-life relationships with godly people" (175). Ouch. That applies to singles and couples alike.
Debra's last chapter called "The Beautiful Exchange" could be a book itself, comparing the seasons of a year to seasons in marriage. I guess it is how you view each natural season how you would compare it to marriage, but it's a challenge to think deeper about the ebb and flow of life seasons.
"Marry someone who will make you better" (184).
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 26, 2018
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