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Description
Rat IL-35 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 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 1000pg/mL). Then dilute to the following concentrations: 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, 15.625pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 1000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 500pg/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. |
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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 an IL-35 capture antibody. After incubation and washing, the sample 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 IL-35 content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat IL-35 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interleukin-35 (IL-35) is an anti-inflammatory cytokine of the IL-12 family. As a member of the IL-12 family, IL-35 is produced by a wide range of regulatory lymphocytes and plays a role in immunosuppression. IL-35 can inhibit the development of Th1 and Th17 cells by limiting early T cell proliferation. IL-35 is a dimeric protein composed of the IL-12α and IL-27β chains, encoded by two separate genes, IL12A and EBI3 (Epstein-Barr virus-induced gene 3), respectively. Its receptors consist of the IL-12Rβ2 (part of the IL-12R) and gp130 (part of the IL-27R) chains. In contrast to these two related interleukins, IL-35 can signal through only one of these chains. This has been demonstrated in vivo, as the absence of either receptor chain does not affect the effects of IL-35. On regulatory B cells, IL-35 signals through the IL-12Rβ2 and IL-27Rα subunits. | |||||||||||||||||||||||||||||||||
| 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 | 15.6-1000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.6 ★★★★★
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Product Reviews
★★★★★ 5
Nice large format printer which is what I needed
Pattern Name: Printer, Style: ET-15000
I make stained glass panels and needed a printer that I could make larger copies with. I also wanted one that I could manipulate my images with. This was my best option and I love the Eco-Tank printers. After reading the reviews I made sure to add a couple years of additional warranty for my peace of mind. Finding 13 x 19 paper that was affordable was a tough challenge. I found a bunch on FB marketplace and bought everything they had to offer.
This printer was very easy to set-up. I just followed the instructions and then followed what was on my computer screen. At first I thought I would have an issue connecting to Wi-Fi but when it failed on the computer, I was able to go to the printer and do it from the control panel - Easy peesey. I have used it several times since filling, charging, and installing and everything works as it should. My husband has an Eco-Tank printer and I've wanted one since he got his. I love that the ink will last a very long time and that it will be cost efficient. I am not overly interested in photo printing so this was the perfect choice for my needs.
The print quality is fairly good when comparing it to my old Canon and e that there are several paper trays and ways to make copies, especially the automatic feed and the larger scanner bed.
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Reviewed in the United States on January 26, 2026
★★★★★ 1
document feeder not working on first real use.
Pattern Name: Printer, Style: ET-15000
Serious reliability issues and concerns. We have owned the printer for about 1 month and printed a limited amount. For the first time today I tried scanning with the document feeder. I was scanning about 12 pages that should of been 1 document. The first page scanned and then stopped. I started the scanner again and it scanned about 5 pages and then jammed. The rest of the pages were a constant jamming/not feeding fiasco. I would run to the computer and hit scan, then run to the printer and manually push on the paper as hard as i could without the paper bending. if I pushed just right i could get a few pages through without jamming. I ended up with about 6 pdf's instead of one document and constant jamming in the paper feeder. Frustrating from a brand new printer and the first time using the paper scanner. There is a clicking noise when it scans. a few of the pages fed properly but the rest had to be forced. No obvious obstructions or other issues. I tried cleaning the rollers with no success. Then attempted to reach out to Epson support. The only support option I could find was an email form that put me in a captia loop after entering all the information so I had to start over on the ticket submission.
No chat support, no phone number, brand new higher priced printer with serious build/reliability issues. Frustrating.
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Reviewed in the United States on April 14, 2026
★★★★★ 5
This is a great printer. I love it.
Pattern Name: Printer, Style: ET-15000
I’ve been using the Epson ET‑15000 for custom chip bags and ran into a huge issue with my original paper choice. The printer itself is great, but the Koala 30 lb glossy paper did not play well with the black ink at all. Even after letting my prints sit for a full two days, the black areas would still smear and come off on my fingers as soon as I started folding and assembling the bags. It was super frustrating and made it hard to trust my prints for orders.
I decided to try Eshang high gloss paper instead, and the difference has been night and day. With the same Epson ET‑15000 and the same designs, the Eshang paper works like a charm. The black ink dries quickly, doesn’t rub off when handled, and holds up beautifully while I’m folding and sealing my chip bags. Colors look vibrant, the finish is nice and glossy, and I don’t have to worry about smudging anymore.
If you’re using an Epson EcoTank like the ET‑15000 and struggling with black ink smearing on Koala glossy, I highly recommend switching to Eshang high gloss. It completely solved the problem for me and made my chip bag printing so much easier and more professional.
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Reviewed in the United States on March 13, 2026
★★★★★ 5
Another great Epson product
Pattern Name: Printer, Style: ET-15000 (Renewed)
Terrific large format printer and doesnt break the bank
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Reviewed in the United States on June 9, 2026
★★★★★ 4
Need scan button to work!
Pattern Name: Printer, Style: ET-15000
Love (almost) everything about this except the scan button. The scan button on the Epson screen does not work. I called Microsoft who tried to fix it through the connection but still didn't work. Sent an e-mail to Epson and no response. Really need this scan button to work so that I can scan to desktop.
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Reviewed in the United States on May 5, 2026