Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Jul 7 - Jul 12
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
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. |
|||||||||||||||||||||||||||||||||
| 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 |
|
|||||||||||||||||||||||||||||||||
| 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. |
|||||||||||||||||||||||||||||||||
| 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 |
Shipping Notes
- Free Standard Shipping on $100+ Orders to the USA.
- Except Preorder products are shipped in 48 hours.
- Delivery to the USA:
- Standard Shipping : 3-10 business days
- If time is of the essence, please consider selecting expedited delivery for faster service.
Exchange/Return Notes
- We offer a 30-day return/exchange service after receiving.
- Final sale items are not eligible for returns or exchanges.
- To process your return/exchange, please contact us at [email protected]
- Please click here for more details>>> Return & Exchange Policy
4.2 ★★★★★
Based on 1512 reviews
Sort
Product Reviews
★★★★★ 5
Best miscellaneous wipe for the money
Scent: Unscented, Size: 25 Count (Pack of 4)
Excellent micellar wipe. Best value for the money.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 16, 2026
★★★★★ 4
Great, but different.
Scent: Unscented, Size: 25 Count (Pack of 4)
I've been buying these for a while now. I love them because I have really sensitive skin and these clean my face without leaving a greasy film, or irritating my skin like other face wipes do. Mind you, the only makeup I wear is mascara though, so it's really only cleaning off oil and dirt. They don't dry out my face either. The only problem I have with these, is that, the last batch I ordered (while in the same packaging, and having no difference in looks or labels) is different than what I had previously been receiving from the same company. They smell different (actually better in my opinion) and they do NOT remove my mascara as well anymore. I still love them, because it does help keep my face from being excessively oily and keeps it clean other than removing my mascara. Though I am disappointed that I now have to get a different brand wipe just for removing my mascara now.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 8, 2020
★★★★★ 5
Best of the facial wipes
Scent: Unscented, Size: 25 Count (Pack of 3)
I have used a lot of facial wipes. But, because of environmental reasons I am trying to cut back on the excess use of wipes which will go into a landfill. However, there are times when I am just too tired or too lazy to do a whole cleansing routine at night and I will reach for a wipe. These do a good job of taking off even waterproof makeup and yet still being gentle on the skin. I find I have no sensitivity to them(and I am sensitive to many, many things). I bought them specifically this time for my trip to Thailand because I knew it would be hot and humid and that I probably would need to cleanse my face a couple of times during the day from sweat and grime. They worked and I found them refreshing. I think I will always keep them on hand for the times I need them — this is by far my favorite brand.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 22, 2022
★★★★★ 5
Great for Rosacea Skincare
Scent: Unscented, Size: 25 Count (Pack of 6)
My elderly mom has rosacea and these moist Micellar Facial Wipes are the absolute best for cleaning her delicate skin. I was so happy to discover Simple Facial Wipes because I had tried many other brands in the past, but all of the other wipes irritated her face and caused a burning sensation. These wipes are soft, sturdy and unscented. They are also pricier than other brands, but they are well worth the cost.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 29, 2025
★★★★★ 5
Best wipes at a great price!
Scent: Unscented, Size: 25 Count (Pack of 6)
These are about the best facial cleansing wipes I've found anywhere, and if you buy the bundle it's a great bargain as well. The wipes are gentle on my sensitive skin, but do a thorough job of getting off the makeup, even mascara and eyeliner. My husband was shaking an ink cartridge the other day and black ink sprayed out all over his face and shirt. On impulse we grabbed one of my facial wipes and started to clean the ink off his face. It easily cleaned up all of the ink, even from his mustache and beard with no residue, and I only had to use two of them. That's how effective these wipes are!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 23, 2020
recommand products
2025 Specialized Rockhopper Mountain Bikes Green
275.00
SCOTT Spunto Kids Mountain Bike Helmet
31.19
2025 Merida eOne-Sixty SL 10K Electric Full Suspension Mountain Bike in Green
3750.00
Mammoth TireBiterII Retriever with Loop Rope Handle – Natural Rubber Dog Toys
15.59
2025 Specialized Tarmac SL7 Sport Road Bike in Black
1124.50