Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Jul 9 - Jul 14
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
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. |
|||||||||||||||||||||||||||||||||
| 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 |
|
|||||||||||||||||||||||||||||||||
| 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. |
|||||||||||||||||||||||||||||||||
| 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 |
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
4.3 ★★★★★
Based on 1281 reviews
Sort
Product Reviews
★★★★★ 5
Great play and chew toy!
Pattern Name: Star
The dogs love these!
They're sturdy and give them plenty of chew time. The size is great and they are very playable. The dogs have a lot of fun and the weight is just enough for our little dogs.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 20, 2026
★★★★★ 5
Great for tug of war
Pattern Name: Checkered
Dog loves them. Best tug of war toy yet. Pretty we’ll indestructible
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 2, 2026
★★★★★ 5
Great Dane Tested and Approved 5 Star
Pattern Name: Checkered, Pattern Name: Checkered
A picture is worth a thousand words applies to this product, as you will see in the attached photos these stand up to the abuse of a Great Dane. He has dragged me around the yard for hours already with this in his mouth and the stretchy handle keeps it shape.
This ball is a little firmer than others of these I have had in the past, which helps exercise his giant jaw. I could get a few weeks out of the other brands I have bought. This brand is still like new well past the time others have failed. I will be ordering more as my French Mastiff is interested in one of his own (the boys do not like to share toys - most dog guardians understand).
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 29, 2024
★★★★★ 4
Holding up well.
Pattern Name: Checkered
Seems to be holding up to my aggressive chewer. I have it hanging from a tree branch and she loves leaping to get it! Buying another pair so she has a greater selection around the backyard.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 3, 2026
★★★★★ 5
Good quality
Pattern Name: Checkered
10/10 my dog lovessss and she chews through everything but they last daily tug of war, I don’t let her sit and chew on it thought I only use it outside
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 28, 2026