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Description
Human LEPROTL1 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. 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 Leptin Receptor Overlapping Transcript Like 1 (LEPROTL1) capture antibody. 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 Leptin Receptor Overlapping Transcript Like 1 (LEPROTL1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Leptin Receptor Overlapping Transcript Like 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Leptin receptor-spiked transcript protein-like 1 (LEPROTL1) is a protein-coding gene associated with diseases such as colorectal adenocarcinoma. An important homologue of this gene is LEPROT. It negatively regulates cell-surface expression of the growth hormone (GH) receptor in the liver and may contribute to hepatic resistance to GH during periods of reduced nutrient availability. | |||||||||||||||||||||||||||||||||
| 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 homogenates and other biological fluids |
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4.1 ★★★★★
Based on 13 reviews
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Product Reviews
★★★★★ 5
Lasts forever
Flavor Name: Bacon, Size: X-Large
Gizmo, my 92lb Labrador puppy, just loves this toy. He’s had one since he was 3 months old, and he just had his birthday.
Absolutely good for hours of contented chewing and tossing.
Highly recommended!
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Reviewed in the United States on June 6, 2026
★★★★★ 4
Our dogs love these bones.
Flavor Name: Bacon, Size: X-Large
ALWAYS SUPERVISE A DOG WHEN THEY ARE CHEWING OR EATING. It doesn't matter what it is, they can choke on anything, we have had multiple instances where we have had to intervene to prevent or stop choking.
I also was going to give this a 5 star review, but after reading the other reviews I didn't. The pet product industry really needs to educate the consumer more about pets and what type would do best with their product. Either way, I think my experience might be helpful to some and we are pretty happy with these bones because the dogs are.
Here's the short version of my recommendations:
I do not recommend Nylabones for dogs that are teething
If an adult dog is chewing non-stop, Kongs with treats or peanut butter are great to help break up the chewing time. Even if they won't chew the Kong, they'll definitely stop whatever they are doing and lick out all of the peanut butter, which is actually calming and tiring for dogs.
It might be worth trying a much larger size bone if you have a smaller dog. I never had small dogs until now, but the large bone seems to tire our 20 pound dogs out quicker than the smaller bones ever did, and their little jaws seem to handle it just fine.
All of our dogs are gnawing with their back teeth at an angle on the white nyla part, as though they are rubbing their teeth against it to clean and sharpen them. The other type of chewing would use more of the front teeth and pull, like they're cleaning meat off of a bone, which isn't the right kind of chewer for Nylabones. Ours only occasionally chew them like that, so I know they are eating some pieces but it doesn't seem to affect their health since it's pretty limited and small (like a crumb).
Here's my experience with these bones:
We got our first Nylabone's for small breed puppies about a 18 months ago (not this product but same brand), and the puppies did try swallowing big chunks, so this type of bone was not great for them yet.
However, our old large breed dog decided he wanted to chew on the small puppy bones too, so I got him the large one of this bone because it's huge with four points, and more his size. It could be a challenge for dogs who don't use their paws like hands, but ours seem to figure it out comfortable ways to chew on it.
The old dog absolutely loved it, and had no big chunk issues. He very quickly figured out not to chew the red part because it is a totally different plastic than the white part (which is whatever "Nyla" is).
My understanding from the package is that the white part is somewhat digestible, or safely passes through in very small pieces. I specifically remember that being why I decided to try these. Our big dog used to choke on raw hides when he was younger, so we had to stop all chewing things (which just meant more treats and walks so he was happy!), but now that he is older he is really enjoying this bone.
Our old dog did wear one down to the nubs after about 3-6 months, and continued chewing on it occasionally (less often since there is less there to chew, and I knew I was supposed to toss it but he loved it so much). So I just got him a new one, but he still carries around the old one to show it off when we come in the door. He's chewing the new one now, so he seems to understand that the old one is 'done', and I haven't felt the need to take it away for safety reasons. The funny part is that the small dogs (now 2 yr old) actually chew on his large bone too, it's pretty comical given the size difference. I got them the extra-small versions and they haven't shown much interest in those.
I decided to also get one for our new 'store' dog who is about 1.5 yrs old and a lab/boxer mix and lives at the store full time. It was to help him transition away from his neurotic tail biting and rope bone that is falling apart from his prior home (where he was EXTREMELY bored and was getting into everything). He has enjoyed the Nylabone for about 2 weeks. I don't think it's worn down too much yet, but he's definitely going to wear his out faster (maybe 2+ months?) than our other three dogs combined, but his teeth are also sharper and he chews more, so that's fair.
So far, no pieces or chunks from the Nylabone. He also hasn't had a problem leaving the red part alone and he's learning how to use his paws more because of the shape of this thing. He still uses it when he's bored, but we've started increasing his training practice, so hopefully he'll be past the chewing phase by 2 yr old, and he has stopped getting into anything (like trash or 'stuff left out' that dogs manage to get into). He's also a huge and a very active dog, so having regular customers and activity help tire him out more than any bone can.
Best of luck!
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Reviewed in the United States on May 31, 2019
★★★★★ 5
Two pits they last forever
Flavor Name: Bacon, Size: X-Small
Dogs love this bone
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 28, 2026
★★★★★ 5
variety of texture and focal points, lab approved
Flavor Name: Bacon, Size: X-Large
My labs dont try to destroy stuff. They just like to gnaw on something once in awhile. This has lasted a very long time. Has the typical dents and other gnaw marks, but has otherwise withstood the test of time. The different texture (very firm but with mild "give") and spots to focus on has made this a favorite go-to toy when the need to gnaw comes about. We often do bully sticks and yak chews for the majority of their chewing needs, but the nylon style also seems to hit the spot.
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Reviewed in the United States on March 8, 2021
★★★★★ 3
I find it sharp edges after just a little bit of chewing
Flavor Name: Bacon, Size: X-Large
My two Labradors are just not interested in this product also it's very sharp after chewing it briefly I like the Unique Style Unfortunately they are not interested maybe it's just them
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Reviewed in the United States on May 17, 2026