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Description
Rat IL-3 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. 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. 5. 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. 6. 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 100 pg/mL). Then dilute to the following concentrations: 100 pg/mL, 50 pg/mL, 25 pg/mL, 12.5 pg/mL, 6.25 pg/mL, 3.125 pg/mL, 1.5625 pg/mL, and 0 pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 100pg/mL standard working solution into the first EP tube and mix thoroughly to make a 50pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube is used 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 utilizes 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 Interleukin 3 (IL-3) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Interleukin 3 (IL-3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Interleukin 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interleukin-3 (IL-3) is an interleukin, a biological signal (cytokine) that, as part of the immune system, can improve the body's natural response to disease. IL-3 is primarily produced by activated T cells to initiate the proliferation of various other immune cell types. However, IL-3 has also been shown to be produced by IgG+ B cells and may be involved in early antibody isotype switching. IL-3 stimulates the differentiation of immature bone marrow mononuclear cells, leading to changes in macrophage and granulocyte populations. IL-3 also induces various effector functions in both immature and mature cells, allowing for more precise regulation of the body's defense against microbial pathogens. IL-3 also participates in platelet remodeling through megakaryocyte development. | |||||||||||||||||||||||||||||||||
| 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 | 1.56-100pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.4 ★★★★★
Based on 2182 reviews
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Product Reviews
★★★★★ 5
It looks really cool and striking.
Color: Grey, Color: Grey
Really cool and beautiful watch. Although my specific unit runs 5 seconds fast every day—at first it’s annoying, but eventually you stop caring. 🤣
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Reviewed in the United States on April 16, 2026
★★★★★ 5
A Beautiful Timepiece at an Awesome Price!!
Color: Grey, Color: Grey
I have lusted after this watch for four months. I first tried on a similar model (stainless steel band with a green face) in a local jewelry shop (Spitz Jewlers, Walnut Creek, CA, USA). It was "love at first sight," even if I didn't love the white lines on the face from 12-6 and 3-9. At the time, the MSRP was $775.
The MSRP has since increased to $795. But I bought this beauty on Amazon.com for only $524.99; after applying the cash-back from my Prime credit card, this watch costs only $498.75. This was "the offer that I could not refuse," to quote from "The Godfather."
Accuracy could be better: after only a few days, it is now 7 seconds behind my phone (to which I set my new watch). But I don't really care about that all that much. This is why I rated "accuracy" at 4 stars, but my overall review is 5 stars.
My video shows the date changing suddenly at 11:59:59pm, just how I want it; in another photo, you can see that my Seiko changes the date more slowly, which I do not like.
This watch is absolutely phenomenally beautiful. If you're on the fence over whether to make this purchase, then I would say "go for it!" You will not find a prettier, more accurate watch at a lower price in ANY store in the entire USA.
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Reviewed in the United States on April 28, 2023
★★★★★ 4
Classy and sporty. Great value!
Color: Grey, Color: Grey
Excellent watch for the price. Looks sporty and dressy. More towards dress watch. Blue is dynamic. Strongly recommend blue for sporty feel. I was thinking of aqua terra. This come close to it with a price less than what i would have paid my state tax for omega aqua terra. My only concern is bracelet which i feel kind of OK. Better bracelet design would have made this watch much superior. 21 lug width is awful. Other than that, nothing to complain.
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Reviewed in the United States on July 13, 2020
★★★★★ 5
Stunning Gentleman's Watch
Color: Grey
This is my second Tissot watch. It's quite stunning in person. The online photos don't do it justice. It has the look and feel of a much more expensive gentleman's watch which makes this purchase a great bargain. Tissot is a great reliable Swiss made watch brand. My first Tissot watch which is probably more than 20 years old still runs great. Very pleased with this purchase.
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Reviewed in the United States on October 16, 2025
★★★★★ 5
Good Tissot Watch
Color: Grey
Arrived on time, in its original box, include a 2 year international warranty, into the box include a book with the Tissot brand history in Switzerland. Algo a second book with a whole catalog of Tissot watches men and women.
The watch looks better when you put in on your wrist. The “naval” blue really gives the clock a classy look. The stainless steel workmanship is really good, the metal is bright and perfectly polished. The metal wristband quality is very good, did not have any problem to adjust the size , I went to my preferred clock workshop for this job, they extracted a few pieces of the band to fit to my small wrist size. Have to get used to the butterfly latch, but after a few days, now I learned to open and close it without any effort. The clock it’s very accurate, I’ve been using my Tissot for near a month and it just increased a few seconds per week. I do the setup and check accuracy with my pc clock. In short, It”s a good value watch, nice, good looking, accurate.
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Reviewed in the United States on August 1, 2021