SKU: 34444038533

Tacrolimus ELISA Kit

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Description

Tacrolimus 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 culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, but avoid repeated freeze-thaw cycles.
Cell lysis buffer: Gently wash adherent cells with pre-chilled PBS, then trypsinize and harvest by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Wash the harvested cells three times with pre-chilled PBS and resuspend in 150-200µL 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.
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 tube. 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 Biotin-Antibody Working Solution: Centrifuge the concentrated Biotin-Antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated Biotin-Antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent) and use on the same day.
4. Preparation of Enzyme Conjugate Working Solution: 15 minutes before use, centrifuge the 100x concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100x concentrated HRP enzyme conjugate to a 1x working concentration with universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Use the same day.
5. Preparation of 1x Wash Solution: Dispense 10mL of 20x Wash Solution into 190mL 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 them to 4°C.
2. Sample Addition: Add 50 μL of sample or standard of varying concentrations to the corresponding wells. Add 50 μL of universal diluent to the blank wells, followed by 50 μL of Biotin-Antibody Working Solution to each well. Cover with film and incubate at 37°C for 1 hour. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate to minimize matrix effects. The sample concentration should be multiplied by the dilution factor when calculating the final concentration. It is recommended to run replicates for all samples and standards.)
3. Plate Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, then shake off the wash solution and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used).
4. Enzyme Conjugate Working Solution: Add 100 μL of enzyme conjugate working solution to each well. Cover with film and incubate at 37°C for 30 minutes.
5. Wash: Discard the liquid and wash the plate five times as in step 3.
6. Add 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.
7. Add 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.
Calculation of experimental results:
Result evaluation:
1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction value. Plot a standard curve for 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 competitive enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled antibody, and HRP conjugate are sequentially added to microwells pre-coated with the universal species Tacrolimus (Tacrolimus) ELISA Kit antigen. After incubation and washing, the assay is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the presence of the universal species Tacrolimus (Tacrolimus) ELISA Kit in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration.
Source All
Synonym General Tacrolimus(Tacrolimus)ELISA kit
Detection Type Competition Law
Composition
Name 9 6 T  match   set remark
Pre-coating 96 Well plate 8 Hole ×12 Strip without
Standard 2 branch
Dilute as per instructions
Universal diluent
2×20mL
without
concentrate Biotin- Antibody ( 100× )
60uL
Dilute as per instructions
Concentrated enzyme conjugate ( 100× )
120uL
Dilute as per instructions
20× Washing liquid
2×10mL
Dilute as per instructions
Bottom thing ( TMB )
10mL
without
Stop liquid
6mL
without
Sealing film
4 Zhang
without
Instructions
1 Share
without
Background Tacrolimus, also known as FK506, is a fermentation product isolated from the genus Streptomyces. It is a macrolide antibiotic and a powerful new immunosuppressant that mainly inhibits the release of interleukin-2 (IL-2) and comprehensively inhibits the action of T lymphocytes. It is 100 times more potent than cyclosporine (CsA).
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 culture supernatant, cell lysate and other biological fluids
Shipping Notes
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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]
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SKU: 34444038533

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4.6 ★★★★★
Based on 21 reviews
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Russell C. Longmire
Cuba, US
★★★★★ 3
Looks Nice But Destroyed by my Lab
Color: Checkers - Elephant (Gray), Size: Large, Color: Checkers - Elephant (Gray), Size: Large
A really cute little elephant. My lab loved it and carried it around for a week. Then the elephant began losing body parts. Both legs are gone now plus the big squeaker. Lab still likes the elephant and carries it around. Its loosing stuffing and the real question is what body part is next. Its nice but not tough.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 26, 2026
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Verified Purchase
Nash Crawford
Whiting, US
★★★★★ 5
Nice toy for Chewers!
Was chewed in a day or so, but she loved it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 13, 2026
B
Verified Purchase
Beth
Bozeman, US
★★★★★ 5
Still in one piece and played with!
Color: Checkers - Dinos Bruto (Purple), Size: Mini
Very durable for my little King Charles Chewer. She's had it for a while now and all though the chewing phase has calmed down, she still can be somewhat rough with the toys. The little dino is standing firm. It was a battle she figured she couldn't win with Dino. Now he's a fetch/ tug o war toy that still gets a few nibbles.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 10, 2026
K
Verified Purchase
Kimmi Bear
Alexandria, US
★★★★★ 4
DONT BUY THE ELEPHANT, ITS NO GOOD. but GR8 BRAND!
Color: Checkers - Elephant (Gray), Size: Large
The elephant toy had no trunk by the FIRST day! The elephant now looks like an angry Koala bear OR an angry trunk less elephant(whichever u want to say) I LOVE the donkey and dinosaur toys so I bought the elephant confidently, thinking I had nothing to worry about. I wouldn't have spent money on this knowing the trunk would be chewed off in a matter of minutes. My Bella usually chews but she has had her donkey and dinosaurs with the chew guard for MONTHS and there are NO TEARS OR RIPS even with her playing and chewing on them ROUGHLY all day. If you buy any of these type toys I DO highly recommend the donkey or dinosaurs ONLY. The elephant being chewed up in a matter of minutes really makes me angry that I ever spent the money on it. HAD I known it wouldn't be the high quality dog toy I'm used to from GoDog, I would've just bought a second donkey or dino! Now Bella doesn't get her new toy for the month (I try 2 buy her a new stuffed toy or 2 about 1 time a month whenever I have the extra money for it) I guess she will have to go without this month because I spent it on this now trunkless elephant/angry koala:( Whichever name you call it, it is now going to be trash. UPDATE! If you contact the GODOG company they will send one replacement toy for your pet! They also asked me my pet's size, breed and age in order to send the best possible toy! They sent Bella a snake that has held up Extremely WELL! It's great and so far is my favorite(and one of Bella's favorites) The snake has less sqeaky things in it so she doesn't have the drsire to tear it apart immediately to retrieve them by rippong it to shreds:) My updated rating is for the MANUFACTURER'S GREAT CUSTOMER SERVICE. The fact that they actually replaced it was good but the fact that they went that extra mile by researching the best toy match for my Bella was above and beyond! If you have a Godog toy that went bust right away, contact the manufacturer for a replacement. If you give them your dog's age, weight and breed they will even try to match your pets replacement toy with something that is better suited to your dog's chewing style. (Bella's style is destructive haha) Good luck, ya'll!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 26, 2016
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Verified Purchase
AmberC
Lake Worth, US
★★★★★ 5
My Dobermans favorite toy to destroy, and one of our recommended picks.
Color: Checkers - Elephant (Gray), Size: Large
Ok, here's an honest breakdown review. I have a 6 month old Doberman who shreds every stuffed animal and dog bed he's come across. He's one of the worst chewers I've owned. We have tried a WIDE variety of stuffed toys. That said, this elephant is his favorite toy. I have ordered 4 of them so far, and just ordered another 4-pack on a buy three get one free deal. The elephant stands out above all the other stuffed creatures from goDog for me as to durability. Hence why I keep buying more. But they usually last about 2-3 weeks on average. In our house, the poor creatures are subjected to vicious, crazy, shaking of the head, helicopter type rotation. It' so hard if it hits your bare arm or leg it actually stings especially when its wet. Then the old, put it between his legs laying down, tearing at the legs action. The limbs are usually the first things to go over time. But they are each almost self sealing, so you clean up as little fluff and keep using it. Eventually the pup reaches the main body and finishes it off. For a normal light to normal chewer, one of two will get you a long way. For those of us resigned to having zoomie type manic chewers that need a constant supply of toys, 2-3 week durability is pretty sound too. I do not recommend the green cheaper monster. It lasted about 45 minutes. The free replacement took about 3 days. Hopefully this aids you in your decision!
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Reviewed in the United States on June 10, 2026

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