Pay in installments of $46.03 with
,
and
Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Aug 24 - Aug 29
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
Description
Rat NETs 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 collected cells three times with ice-cold PBS and resuspend 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 remove the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 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. |
|||||||||||||||||||||||||||||||||
| 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 a neutrophil extracellular trap network (NETs) 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 neutrophil extracellular trap network (NETs) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Neutrophil extracellular trap network ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
|
|||||||||||||||||||||||||||||||||
| Background | Neutrophil extracellular traps (NETs) are primarily composed of neutrophil DNA, which binds to pathogens. Neutrophils are the immune system's first line of defense against infection and are traditionally thought to kill invading pathogens through two strategies: phagocytosis of microorganisms and secretion of antimicrobial substances. NETs enable neutrophils to kill extracellular pathogens while minimizing damage to host cells. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant 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
4.8 ★★★★★
Based on 22 reviews
Sort
Product Reviews
★★★★★ 3
Good But
Format: Kindle
A little disjointed at times, at least to me. This was about Lila (omega woman) and Harrison, Wesley and River (alpha males). The main characters were likeable, as was Lila's friend, Wendy. Lila finally wised up and left the alpha she had been with for a couple of years because he was an a-hole, who treated and used her horribly. She knew some, if not all, of the guys from college if I remember correctly. The guys had been looking for an omega to bond with for some time. When Lila saw Harrison again, the other two guys were out on a date with another omega but Harrison refused to go because he didn't like her. He saw Lila again and knew she was the omega they had been searching for. However, it wasn't easy for them to convince Lila of that because of the way her ex-alpha treated her.
The sex was hot. There was one inconsistency. I could almost swear that when Lila first saw Harrison again, she told her best friend, Wendy, that Harrison was the lead for his pack. Then later on when she's spending time with River, she thinks he is the pack's lead. Maybe I read something incorrectly. All in all, a good story.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 13, 2026
★★★★★ 5
Sweet read
Format: Kindle
A sweet steamy read of an omega who finds her worth and finds the guys that are willing to show her. The story had lovable characters with low angst and a HEA.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 7, 2025
★★★★★ 5
knotty book 1
Format: Kindle
Super enjoyable! What a horrible ex. Was glad to see the back of him, I was hoping I would get to see him suffer a whole lot more.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 5, 2025
★★★★★ 5
Short and sweet
Format: Kindle
Beautifully written, I was worried when it was labelled as a novella at the start, really used to those missing info and jumping forward to complete the story or not having complete character growth, not to worry! Author gave enough time to fall in love (and hate) with everyone, can't wait to read Wendy's story I feel like she will be the badass we all love!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 17, 2025
★★★★★ 3
Likeable characters, but only 75% complete story
Format: Kindle
I really liked the characters introduced, but the author didn’t fully conceptualize everything. We know Lila works long hours, but despite being a college graduate we never learn anything about what she does, where she works, how much she gets paid, etc. For an omega who seemed to be able to support herself, that seemed an unusual oversight. We learn absolutely nothing about what she wants in life, either. Is she just surviving the post-college life struggle or working up to her dream job? Fiona the flamingo had more detail than some of the main characters, but she was 10/10 as good as Wendy for a side character.
— spoilers ahead —-
Separately, some of the plot details were contradictory, or just missing. On one page Lila and Larry were together for 18 months, on another it was 4 years. When did she even move in with him? What did she do while he was gaming? How long were she and Wendy friends for Lila to trust her like that but not know much about her? And why didn’t Lila’s alphas sue Larry to reduce Lila’s debt? Shopping seemed a weird choice pamper her without knowing more about Lila’s debt and plans to reduce it. And her alphas never even ask anything about her job, but she’s The One. Such a frustrating relationship plot hole.
We never even learn anything about the alphas’ families or what they want in life either. I wish the author had written about 30 more pages fleshing out the details and showing how the alphas were just as in love with Lila’s personality as they were with her scent and body. Loved the concept, but I still feel like I read 3/4 of a book.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 29, 2025
recommand products
LA 835 - County Lunatic Asylum, Lancaster, Lancashire
10.95
Restoration Two-Handle 3-Hole Deck Mount Bridge Kitchen Faucet
191.74
Milano Three-Handle 5-Hole Deck Mount Roman Tub Faucet with Hand Shower
333.11
LA 2008 - W & W Johnson, 85 Bradshaw Gate, Delivery Lorry, Bolton, Lancashire
13.00
Wilshire Two-Handle 3-Hole Deck Mount Roman Tub Faucet
216.11