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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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4.3 ★★★★★
Based on 1708 reviews
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Product Reviews
★★★★★ 5
A Hilarious, Action-Packed Thrill Ride!
Format: Hardcover, Format: Hardcover
A Coast Guard veteran and all around good guy, Carl, lives in an apartment with his ex-girlfriend’s cat, Princess Donut. On a cold, winter night, Donut slips out of a window and gets stuck up a tree. And it’s a good thing she does! Wearing nothing but a jacket, boxers, and a pair of slippers that don’t fit, Carl goes outside to try to coax her down. He’s just about retrieved the cat, and then it happens.
The whole world is changed. In the blink of an eye, every building, car, and piece of technology on the planet is flattened. Smooshed. Gone. A bodiless voice announces that anyone who doesn’t want to live off whatever is left on the planet will need to enter stairs. Carl and the cat do so, and that’s when the fun starts.
It seems the galaxy has had a long-running and massively popular television program that follows “dungeon crawls”—classic role-playing/video game scenarios where adventurers go into a medieval dungeon, explore, fight monsters, win treasure, gain experience, become more powerful, and then proceed to deeper, harder levels. Earth has been selected to serve as the setting for the current season. That’s right. The Earth has been destroyed for the sake of a galactic television game. By entering the stairwell, Carl, Donut, and a couple million other humans have become participants in this game. Instead of remaining a pet, Donut is made into a fellow “crawler,” like Carl. She can speak, and reason, and fight—all with the personality one would expect from a cat named Princess Donut
The rules to this galactically televised dungeon crawl are intricate. But essentially, Carl and Donut begin to mentally see stat screens, just like in an RPG video game: health, various skills, their strength, dexterity, intelligence, and constitution. In classic 80’s kids Dungeons & Dragons style, they have unlimited encumbrance, meaning they can carry anything they can pick up, file it away in “inventory,” and pull it up whenever needed. They‘re on level 1 of this season’s crawl, a classic dungeon with tunnels, doors, chambers, and monsters—lots of different monsters. There’s a countdown running, so they only have so many days to find a set of stairs that will lead them down to the next, harder level. And if they don’t find the stairs before the timer runs out, the level they’re on will collapse. There’s all sorts of lethal dangers awaiting the crawlers. And that’s what takes up the bulk of the book.
There are daring encounters, puzzles to sort through, and lots and lots of monsters to fight. In each encounter, the reader is given real time stats of the characters. After their initial shock, Carl and Donut slowly form an endearing partnership, one that proves quite successful in this dangerous game they‘re forced to play.
I’ll confess for the first quarter of the book, I was skeptical. It felt an awful lot like one of my kids watching someone else playing a video game (which is something I don’t really understand). But Matt Dinniman does a masterful job of weaving in enough subplots—both inside and outside the dungeon—so that both a cohesive story and genuine character development emerge from all the excitement of fighting kobolds, or rigging goblin explosives, or figuring out how to slay a “big boss” monster that vaguely resembles a cat-hoarding old lady. There’s depth to this dungeon.
And of course there’s action. It’s compelling, page-turning, fun. And funny. Dinniman has a sharp, occasionally crass, often dark sense of humor and he knows how to use it in all the right places. There’s snark, and absurdity, and physical comedy, and some snort-through-your nostrils lines. Think of a homebrew Dungeons and Dragons campaign melded with a Hitchhiker’s Guide to the Galaxy vibe that‘s centered around a likable hero and a hilariously self-absorbed cat.
I thoroughly enjoyed this book, and will definitely be pursuing the series. Highly recommended.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 27, 2025
★★★★★ 5
Wish I’d jumped in sooner
Format: Kindle
Great book, easiest read I’ve had in years. Particularly enjoyed switching between the audio and reading. One of the best audiobooks out there for sure, not quite a radio play but the characters do all get proper voice acting and they are brilliant. Book one had me hooked but book 2 really sealed the deal, grateful that there are so many more to read. The comedy, the horror, the bonkers world building, and some really great character work make one of the most insane setups for a book feel easy to buy into and believe in. I’ve been looking for a series that captures my imagination like this for a while and I think I’ve found it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 9, 2026
★★★★★ 5
Funny, smart and nerdy
Format: Kindle
Are you now or have you ever been a member of a TTRPG group or serious video gamer? This book is for you. You'll get all the in-jokes, understand the process and enjoy the story. It's almost literally a step-by-step description of a dungeon crawl from hell, but I was never bored. Matt Dinniman's tone and how he writes Carl are smart and enjoyable.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 30, 2026
★★★★★ 4
Chaotic & absurdly funny!
Format: Paperback, Format: Paperback
Dungeon Crawler Carl was one of those books where the first thought in my head was, what on earth am I reading? And somehow that’s exactly why it works. It’s chaotic, absurdly funny, and completely outside the usual genres I gravitate toward, but it turned out to be such a fun ride.
The premise alone is wild. Earth collapses into a giant dungeon run as a galactic game show, and Carl ends up fighting through it alongside his ex-girlfriend’s cat, Princess Donut, who honestly steals the show for me. Like if I ever get a cat I will probably named her Princess Donut haha! The whole thing is nonstop action, monsters, traps, loot drops, and ridiculous commentary about survival being tied to entertainment value. It’s very LitRPG, very Dungeons & Dragons energy, and packed with pop culture references.
Did a hybrid read and listened to the audiobook when on the go, which is phenomenal and probably the best way to consume it. The narration makes the humor and chaos land even harder. Carl and Princess Donut as a duo are hilarious, and I can already tell this is the kind of series I’ll return to whenever I need a break from heavier reads.
It’s intense, bizarre, and honestly kind of addictive, not something I would jump back to back considering there are like 9 other books, but it is a surprisingly great palate cleanser.
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Reviewed in the United States on March 18, 2026
★★★★★ 5
hilarious, fun and fantastic writing
Format: Kindle
It’s been a little while since I laughed hysterically from a book.
The writing, top tier, the humor immaculate and the characters, compelling.
The story is a mix of satire humor and all around packed with all the things that make a book fantastic. Intrigue, mystery, actual thought. 🤣
For all my booktok girlies who are on the fence, just do it. It scratches an itch I cannot describe.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 22, 2026
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