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Description
Human LGMN 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. 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 the collected cells three times with ice-cold PBS and resuspend them 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 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 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 utilizes 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 legumain (LGMN) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of legumain (LGMN) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Legumain ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Asparagine endopeptidase (LGMN) is a proteolytic enzyme from the C13 peptidase family that utilizes the thiol group of cysteine residues as a nucleophile to hydrolyze peptide bonds (hence, it is also called a cysteine protease). It is also known as citvac, proteinase B, hemoglobinase, PRSC1 gene product, vicilin peptide hydrogenase, and legume endopeptidase. It is encoded by the LGMN gene (formerly symbolized as PRSC1). It hydrolyzes substrates C-terminal to asparagine residues. It can be detected in spleen, liver, brain, testicular tissue, and heart, and is primarily localized in lysosomes and endosomes. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates, cell lysates and other biological fluids |
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4.6 ★★★★★
Based on 505 reviews
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Product Reviews
★★★★★ 3
Interesting take on the genre
Format: Kindle
True rating: 3.25 ⭐️
I enjoyed the fresh take on the genre. The best way I could describe the setting and world is an apocalyptic dystopian version of Farie where vampires, fae, and angles struggle to survive in what is left of the world. It was definitely interesting throwing the academy/hunger games aspect into this world as well.
Even though I guessed the final reveal early on in the book, I kept hoping I was wrong, and it would take a surprising turn. While the "plot twists" were a bit predictable to me, I still enjoyed the ride this book took me on.
Another downfall for me was the plot holes in the world building... I.E. if society has fallen and the world is in the aftermath of war, how are there trains running around the world? Just to take young adults to the trials to get into the golden city? How is the train maintained, the tracks clear, etc?
However, I did enjoy the FMC & MMC and thought they were fleshed out nicely. I also enjoyed the side characters but wish some were developed more like Ashalin (sp?). I do find myself rooting for the MCs to succeed and find happiness together, which is obviously an important aspect for romantasy.
Overall, was this an earth-shattering, mind-bending, terrific piece of literature? No. But was it the worst thing I've read this year? Also, no. This book has, to me, the bones of a great read & just needs a bit more to push it from an alright book to a great book.
Overall ratings:
Plot- 3.5⭐️
World building 3⭐️
Spice 2.5 🌶🌶
Main characters 4 ⭐️
Supporting characters 3.5⭐️
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 12, 2024
★★★★★ 5
great book
Format: Kindle
I am really excited to meet the author at the book retreat this month. I really enjoyed this world that she built and most of the female main character Huntress is so awesome. She goes through a lot in this book and the ending; wow! I wouldn't have even guessed. I highly recommend everyone to read this book.
I have been so lucky this year that almost all the books I have read have been, so far, 5 out 5 stars.
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Reviewed in the United States on June 2, 2026
★★★★★ 4
Fallen for the Fallen Angel – A Guilty Pleasure Worth Every Page
Format: Kindle
There’s something deeply irresistible about a dark academia or trial-based setting, a brooding and arrogant fallen angel, and a fierce heroine with enough sass to go toe-to-toe with him. Wings So Wicked is exactly that kind of book—and I devoured it in just a couple of days.
To be fair, the plot isn’t groundbreaking. If you’re looking for something fresh and innovative in terms of storyline, this might not be it. But if your reader heart beats faster at the mere mention of enemies-to-lovers, jealousy-fueled banter, magical trials, betrayals, and forbidden tension—you’ll feel right at home. It’s like catnip for those of us with this particular weakness.
The chemistry between the leads could have used a slightly slower burn to make the tension sizzle longer, but I still found myself completely invested in their dynamic. There are moments and phrases that feel a bit cheesy or underdeveloped, but honestly? I didn’t care. The vibes were exactly what I wanted.
This book isn’t trying to reinvent the genre—it’s here to give readers like me what we crave: high-stakes magical drama, angsty romance, and the thrill of watching a badass girl and her brooding counterpart clash and spark. If that sounds like your kind of story, Wings So Wicked will hit the mark.
Here’s hoping Book 2 turns up the heat and keeps the magic alive.
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Reviewed in the United States on May 20, 2025
★★★★★ 5
my new favorite book
Format: Kindle
Ok so I never write reviews but this book was so good I felt the need to write this. Firstly your introduced to Huntyr you see her closed off hard core badass than towards the end you see the most subtle change and growth it’s amazing and the enemies to friends to lovers was just perfect, AND THE TWIST AT THE END GOT ME GOOD! You see one spicy scene the whole book but it doesn’t even MATTER BECAUSE THE BOOK WAS THAT GOOD. I’ve read 85 books in 2023-2024 so far and I’m pround to say this is my all time favorite. I’m so excited to read more of Emily Blackwoods books, this was my first time reading one of hers and I’m glad I did because HOLY!! Well done Emily well done
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Reviewed in the United States on February 23, 2024
★★★★★ 4
Fast paced romantasy you will not want to put down!
Format: Kindle
4.25 stars! I LOVED this book with similar vibes to Hush Hush, Fourth Wing, and The Serpent and the Wings of Night! It was fast paced with easy world building and will keep you turning the pages late into the night because you will not want to put it down!
Huntyr is a fierce bad@ss FMC trained to kill vampyres her entire life. She is sent on a mission to go to the academy and earn her spot into The Golden City. Upon arrival, she is forced to room with the delicious fallen angel, Wolf, who is the only one who knows about her assassin identity. The romance, the plot twists, the secrets revealed, the battles, and the tantalizing training scenes had me hooked! And that ending…. I’m holding my breath in need to know hell!
Read if you love:
🪽 Fae, Vampyres, Fallen Angels
🪽 Academy setting with magical trials
🪽 Forced proximity and slow burn
🪽 Rivals to lovers
🪽 Hidden identities and secrets
🪽 Tend your wounds
“𝘖𝘧 𝘤𝘰𝘶𝘳𝘴𝘦 𝘐 𝘸𝘢𝘴 𝘸𝘢𝘵𝘤𝘩𝘪𝘯𝘨 𝘺𝘰𝘶. 𝘐 𝘤𝘰𝘶𝘭𝘥𝘯’𝘵 𝘭𝘰𝘰𝘬 𝘢𝘸𝘢𝘺 𝘧𝘰𝘳 𝘢 𝘮𝘰𝘮𝘦𝘯𝘵, 𝘦𝘷𝘦𝘯 𝘪𝘧 𝘐 𝘵𝘳𝘪𝘦𝘥.”
“𝘐𝘧 𝘺𝘰𝘶 𝘸𝘢𝘯𝘵 𝘮𝘦 𝘰𝘯 𝘮𝘺 𝘬𝘯𝘦𝘦𝘴, 𝘏𝘶𝘯𝘵𝘳𝘦𝘴𝘴, 𝘢𝘭𝘭 𝘺𝘰𝘶 𝘩𝘢𝘷𝘦 𝘵𝘰 𝘥𝘰 𝘪𝘴 𝘢𝘴𝘬.”
“𝘠𝘰𝘶 𝘥𝘰 𝘯𝘰𝘵 𝘬𝘯𝘰𝘸 𝘵𝘩𝘦 𝘷𝘪𝘰𝘭𝘦𝘯𝘤𝘦 𝘵𝘩𝘢𝘵 𝘳𝘶𝘯𝘴 𝘵𝘩𝘳𝘰𝘶𝘨𝘩 𝘮𝘺 𝘷𝘦𝘪𝘯𝘴, 𝘣𝘦𝘨𝘨𝘪𝘯𝘨 𝘮𝘦 𝘵𝘰 𝘰𝘣𝘭𝘪𝘵𝘦𝘳𝘢𝘵𝘦 𝘢𝘯𝘺𝘰𝘯𝘦 𝘸𝘩𝘰 𝘭𝘢𝘺𝘴 𝘢 𝘩𝘢𝘯𝘥 𝘰𝘯 𝘺𝘰𝘶.”
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Reviewed in the United States on June 12, 2024