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Description
Human E-selectin ELISA KitProduct Specification Usage Self prepared test equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision sampler and gun tip: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 incubator 4. Distilled water or deionized water Sample handling and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or 4 overnight, then centrifuge at 1000 g for 20 minutes, take the
Product Specification
| Usage |
Self-prepared test equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision sampler and gun tip: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37 ℃ incubator 4. Distilled water or deionized water Sample handling and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or 4 ℃ overnight, then centrifuge at 1000 × g for 20 minutes, take the supernatant, or store the supernatant at-20 ℃ or-80 ℃, but avoid repeated freezing and thawing. Plasma: Collect specimens with EDTA or heparin as anticoagulants, and centrifuge the specimens at 2-8 ℃ 1000 × g for 15 minutes within 30 minutes after collection. Take the supernatant for detection, or place the supernatant at-20 ℃ or-80 ℃ for storage, but avoid repeated freezing and thawing. Tissue homogenate: Wash the tissue with pre-cooled PBS (0.01 M, pH = 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results), and break the tissue into pieces after weighing. Combine the crushed tissue with the corresponding volume of PBS (generally according to the weight-to-volume ratio of 1: 9, for example, 1g of tissue sample corresponds to 9mL of PBS. The specific volume can be appropriately adjusted according to the needs of the experiment and recorded. It is recommended to add protease inhibitor to PBS) add to a glass homogenizer and grind thoroughly on ice. For further lysis of tissue cells, the homogenate can be sonicated, or freeze-thawed repeatedly. Finally, the homogenate was centrifuged at 5000 × g for 5-10 minutes, and the supernatant was taken for detection. Cell lysate: Adherent cells were gently washed with pre-cooled PBS, then digested with trypsin, centrifuged at 1000 × g for 5 minutes and collected; The suspended cells can be collected directly by centrifugation. The collected cells were washed 3 times with pre-cooled PBS, and 150-200uL PBS was added to every 1 × 10 ^ 6 cells to resuspend (it is recommended to add protease inhibitors to PBS; if the content is very low, the PBS volume can be appropriately reduced) and the cells were disrupted by repeated freezing and thawing or ultrasound. The extract was centrifuged at 2-8 °C at 1500 × g for 10 minutes, and the supernatant was taken for detection. Cell culture supernatant: Please centrifuge at 1000 × g for 20 minutes, take the supernatant for detection, or store the supernatant at-20 ℃ or-80 ℃, but avoid repeated freezing and thawing. Other biological liquids: centrifuge at 1000xg for 20 minutes, and take the supernatant for detection. Preparations before testing: 1. Please take out the kit from the refrigerator 10 minutes in advance and balance it to room temperature. 2. Preparation of standard gradient working solution: Add 1mL of universal diluent to the freeze-dried standard, let it stand for 15 minutes until it is completely dissolved, and then gently mix it (the concentration is 10ng/mL), and then dilute according to the following concentrations: 10ng/mL, 5ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, 0ng/mL. Double dilution method: Take 7 EP tubes, add 500uL of universal diluent to each tube, draw 500uL of 10ng/mL standard working solution into the first EP tube and mix well to prepare 5ng/mL standard working solution. According to this step, suck and mix well in sequence. The last tube is directly used as a blank hole, so there is no need to suck liquid from the penultimate tube, as shown in the figure below. 3. Preparation of biotinylated antibody detection working solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000 × g for 1 minute, and dilute 100 × concentrated biotinylated antibody to 1 × working concentration with universal diluent (example: 10uL concentrated solution + 990uL universal diluent), now prepared for use. 4. Preparation of enzyme conjugate working solution: Centrifuge 100 × concentrated enzyme conjugate at 1000 × g for 1 minute 15 minutes before use, and dilute 100 × concentrated HRP enzyme conjugate to 1 × working concentration with universal diluent (example: 10uL concentrated solution + 990uL universal diluent), now prepared for use. 5. Preparation of 1 × washing liquid: Take 10mL of 20 × washing liquid into 190mL distilled water (the concentrated washing liquid taken out of the refrigerator may have crystals, which is a normal phenomenon. It can be placed at room temperature and then prepared after the crystals are completely dissolved). Operation steps: 1. Take out the required slats from the aluminum foil bag after equilibration at room temperature for 10 minutes, and seal the remaining slats with a ziplock bag and put them back to 4 °C. 2. Add samples: Add samples or standards of different concentrations to the corresponding wells at 100uL per well, and add 100uL of universal diluent to the blank wells. Incubate at 37 °C for 60 minutes after covering the plate sealing film. (Recommendation: Dilute the sample to be tested with a universal diluent at least 1 times and then add it to the enzyme labeled plate for testing. This reduces the influence of matrix effect on the test results. Finally, the sample concentration needs to be multiplied by the corresponding dilution factor when calculating. It is recommended to set up double wells for all samples and standards to be tested during testing). 3. Add biotinylated antibody: take out the enzyme labeled plate, discard the liquid, and do not wash it. 100 uL of biotinylated antibody working solution was directly added to each well, and the plate sealing membrane was covered and incubated at 37 °C for 60 minutes. 4. Plate washing: Discard the liquid, add 300uL 1x washing liquid to each hole, let it stand for 1 minute, throw away the washing liquid, pat dry on absorbent paper, and repeat washing the plate 3 times (you can also use a plate washing machine to wash the plate). 5. Add enzyme conjugate working solution: Add 100uL of enzyme conjugate working solution to each well, cover the sealing membrane and incubate at 37 °C for 30 minutes. 6. Wash the plate: Discard the liquid and wash the plate 5 times according to the washing method in step 4. 7. Add substrate: Add 90uL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37 °C in the dark for 15 minutes. 8. Add stop solution: Take out the enzyme plate, directly add 50uL of stop solution to each well, and immediately measure the OD value of each well at a wavelength of 450nm. Calculation of experimental results: Result judgment: 1. Calculate the average OD value of the standard product and the sample double well and subtract the OD value of the blank well as the correction value. Taking concentration as abscissa and OD value as ordinate, the standard curve of four-parameter logic function is drawn on double logarithmic coordinate paper. 2. If the OD value of the sample is higher than the upper limit of the standard curve, it should be properly diluted and retested and multiplied by the corresponding dilution factor when calculating the sample concentration. |
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| Theory | This kit uses double antibody sandwich enzyme-linked immunosorbent assay (ELISA). To the microwells pre-coated with E-selectin capture antibody, sample, standard product, biotin-labeled detection antibody, and HRP enzyme conjugate were added sequentially, incubated and washed in the middle, and colored with substrate TMB, which was converted into blue under the catalysis of peroxidase (HRP) and final yellow under the action of acid. There is a positive correlation between the depth of color and the E-selectin in the sample. The absorbance (0D value) was measured with a microplate reader at a wavelength of 450nm, and the sample concentration was calculated. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human E-selectin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | E-selectin, also known as CD62 antigen-like family member E (CD62E), endothelial cell-leukocyte adhesion molecule 1 (ELAM-1) or leukocyte-endothelial cell adhesion molecule 2 (LECAM2), is a selectin cell adhesion molecule expressed only on endothelial cells activated by cytokines. Like other selectin, it plays an important role in inflammation. E-selectin is encoded by the SELE gene. It has a cassette structure: an N-terminal C-type lectin domain, an EGF (epidermal growth factor)-like domain, 6 sushi domain (SCR repeat) units, a transmembrane domain (TM), and an intracytoplasmic tail (cyto). | |||||||||||||||||||||||||||||||||
| General Notes | 1. Carry out incubation in strict accordance with the specified time and temperature to ensure accurate results. All reagents must reach room temperature 20-25 °C prior to use. Store reagents in refrigeration immediately after use. 2. Incorrect plate washing may lead to inaccurate results. Make sure to drain the liquid from the wells as much as possible before adding the substrate. Do not allow the wells to dry out during incubation. 3. Eliminate the residual liquid and fingerprints at the bottom of the plate, otherwise it will affect the OD value. 4. The substrate color development solution should be colorless or very light in color, and the substrate solution that has turned blue cannot be used. 5. Avoid cross-contamination of reagents and specimens to avoid wrong results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Any reaction reagent cannot come into contact with the bleaching solvent or the strong gas emitted by the bleaching solvent. Any bleaching component will destroy the biological activity of the reaction reagents in the kit. 8. Expired products cannot be used, and components with different item numbers and batch numbers cannot be mixed. 9. Recombinant proteins from sources other than the kit may not match the antibodies in this kit and are not recognized. 10. If the disease may be spread, all samples should be managed well, and the samples and testing devices should be handled according to the prescribed procedures. |
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| Storage Temp. | Unopened kit, stored at 4 °C, shelf life 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.15-10ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates, cell lysates, cell culture supernatants and other biological fluids |
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4.1 ★★★★★
Based on 6 reviews
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Product Reviews
★★★★★ 5
Great entertainment, charges quickly!
Color: Orange
Super fun entertainment for dog! Toy ball charges quickly and the charge lasts for at least an hour of playtime for the dog. There are 2 modes. one mode Just lights up, which makes it easy for pet to chase it. The second mode not only lights up, but it also wiggles on the ground Randomly and speeds off in a direction on its own, which really makes it interesting for your dog or cat to follow! Once the ball is not touched for about thirty seconds, it goes dormant and it will stay like that As long as it's charged until somebody touches it, and then it starts up randomly lighting up and moving around the floor on its own And making a vibration noise. It's a lot of fun to watch the dog chase and play with, and it certainly keeps them entertained for a while! The laughs and fun entertainment are definitely worth it. The only downside is that my dog chewed off the soft fabric covering in about twenty minutes.And she's not a real big chewer. It looks better with the covering on it, but even without it, the ball works fine and she plays with it every day.So that is not a showstopper. But good to supervise your dog for safety.In case they are a chewer, and they decide to destroy the cover. But the ball works fine without the cover too!
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Reviewed in the United States on March 12, 2026
★★★★★ 4
Looooves her crazy new ball
Color: Orange, Color: Orange
Our 90lb mastiff/boxer mix Indie is very playful, curious and energetic. She needs toys that engage her and keep her busy. This ball does the job!
We’ve had it for less than a week. Here’s what I love and what could be better:
PROS:
• She dog loves it. She whines for it. She gets absurdly excited when I get it out. She’s a bit of an anxious dog, but had zero problems with it inexplicably coming to life.
• It lasts a while on a single charge, so it keeps her busy for a good long time. The longest we’ve used it at a stretch is an hour.
• It’s pretty durable. She’s a super chewer (destroyed every “non-destructible” toy we’ve purchased for her), but the actual ball is hard plastic. She gnaws on it and it gets scratches, for sure. But she’s not going to chew it to pieces.
• On carpet, it’s reasonably quiet.
• It comes with a replaceable silicone button.
• It’s easy to charge…just pull off the silicone button and the charging port is inside.
• The silicone button is flush with the ball’s hard plastic surface, so there’s no way for her to pull the silicone button out and swallow it or get to the charging port.
CONS:
• Cover is worthless if your dog is a chewer. It goes on like an envelope with a Velcro closure. We took it off immediately.
• Without the cover, it’s very noisy on hardwood floors. To us, it’s worth it. (Though, I work from home, so I might feel differently after the holiday vacation.)
• She’s an XL dog, so she can fit the whole ball in her mouth easily and accidentally chomp down on the button and turn the ball off. Not a big deal, just something to know if you’ve got a Lab or larger dog.
• With extended playtime, the two halves can come unscrewed, especially with a large dog who uses their paws/jaws on it a lot. Thankfully, it makes a loud buzzing noise when this happens, so we know we need to take it from her and screw it back together. Is this a potential danger? With the rechargeable battery inside, I’d say yes. But we also don’t let her play with this toy without supervision. So keep that in mind.
Bottom line:
It has its drawbacks, but it keeps her entertained and seems like it should last her at least 6 months with supervised play. Which is honestly REALLY good when we’re talking about Indie, Destroyer of Toys. I’d buy it again.
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Reviewed in the United States on December 31, 2024
★★★★★ 3
Difficult/impossible to unscrew plastic cups (in order to recharge the vibrating/blinking device)
Color: Orange
Generally, I don't leave feedback on items purchased. However, in this case, I feel compelled to do so in hopes the mechanical engineers (of this product) will take my input to heart and ultimately modify it.
First the pros...
- I have 2 high-energy dogs (Jack Russell Terriers) who love their toys (e.g., chewing bones, tug-of-war, etc.) but usually get bored with them rather quickly.
- This interactive PetDroid ball, however, keeps them busy for a long time.
- In fact, my smaller dog (Max) knows its storage location and sits (as early as 6 AM) in front of the cabinet so that I give him "his ball". It keeps him busy for up to 1-2 hours at a time.
- Max is literally addicted to the PetDroid ball... his determinant play with it makes me laugh all the time.
Now, the cons...
- After several days of play, the exterior looks chewed up quite a bit. That's no problem though.
- Unfortunately, the interior "threads/grooves" of the two plastic cups (for closing/fasting) are very tiny.
- Thus, after an hour-long hard play, it is almost impossible to unscrew the 2 plastic cups in order to recharge the interior device.
- It appears the 2 cups somehow come off track and get realigned from the inside.
- At that point, I cannot unscrew the PetDroid ball in order to recharge the device.... and without the blinking/vibration, this toy is no longer fun for play.
- Ultimately, I had to place the bottom half of the ball into a vise and then use a plumbing wrench to untwist it.
Proposed Solution:
- Increase the size of the plastic cups' interior threads/grooves so that they won't get realigned/come off track.
- This would mitigate the current dilemma of NOT being able to unscrew the ball in order to recharge it.
In my view, this should be a relatively simple fix and significantly improve the durability of this product.
Thank you for your time.
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Reviewed in the United States on February 4, 2022
★★★★★ 5
Good fun and entertaining!
Color: Orange
A really fun interactive toy for your dog! It's pretty erratic and really gets him chasing it. The included soft cover was kinda hard to install and then it's hard to turn it off and on with the cover on, I feel like the cover didn't really improve it at all, sure it's louder without the cover but I feel like dog also likes it that way, he likes loud squeaky toys and this thing making a bunch of noise is appealing to him. We have mostly hardwood floors so it really gets moving, I'm sure on carpet it would be a bit quieter and maybe even a bit slower to move around but on hard smooth surfaces this things can really get zipping around abd get some speed to make him chase it. Overall a fun and entertaining toy.
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Reviewed in the United States on January 27, 2026
★★★★★ 5
Awesome
Color: Orange
Awesome Toy . My Jack is crazy over it. She usually get bored with others, but this toy still has her attention after a couple of weeks Runs a long time on a short charge. 👍🏻
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Reviewed in the United States on May 30, 2026