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Description
Rat MIF 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP enzyme conjugate are sequentially added to microwells pre-coated with a Macrophage Migration Inhibitory Factor (MIF) 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 Macrophage Migration Inhibitory Factor (MIF) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Macrophage Migration Inhibitory Factor ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Macrophage migration inhibitory factor (MIF), also known as glycosylation inhibitory factor (GIF), L-dopa isomerase, or phenylpyruvate isomerase, is a protein encoded by the MIF gene. This gene encodes a lymphokine involved in cell-mediated immunity, immunoregulation, and inflammation. MIF regulates macrophage function in host defense by inhibiting the anti-inflammatory effects of glucocorticoids. This lymphokine forms a complex with the JAB1 protein near the cell membrane periphery. MIF is a key regulator of innate immunity. The MIF protein superfamily includes a second member with relevant functional properties, D-dopa isomerase (D-DT). CD74 is the surface receptor for MIF. Macrophage migration inhibitory factor assembles as a trimer composed of three identical subunits. Each monomer contains two antiparallel α-helices and a four-stranded β-sheet. These monomers are arranged around a central channel with three-fold rotational symmetry. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.8 ★★★★★
Based on 21 reviews
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Product Reviews
★★★★★ 3
Not bad
Flavor Name: Superior Blend, Size: 1.4 Ounce (Pack of 1)
Great if you like a more subtle flavor, not bitter. If you just add it to milk then it has a slight very brief strong aftertaste, but I like mine a bit stronger. it mixed well it did not clump for me, it was a great vivid color. I will say I prefer a strong, bold matcha taste in my lattes so I do not think I would purchase again but overall it is a great option for beginners, people trying to figure out what they like, or folks who like a smoother, not bitter taste. The tin did come dented unfortunately, not a huge deal but also not great to receive a product that has been somewhat damaged.
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Reviewed in the United States on April 16, 2026
★★★★★ 5
Creamy, Smooth, and Fresh — Perfect First Matcha Experience
Flavor Name: Superior Blend, Size: 1.4 Ounce (Pack of 1), Flavor Name: Superior Blend, Size: 1.4 Ounce (Pack of 1)
I tried the Naoki Superior Blend Matcha for the first time and was honestly impressed from day one. I prepared it with milk, and the matcha turned out incredibly creamy and smooth with a natural sweetness — no bitterness at all.
The color was a vibrant bright green, which immediately made it feel fresh and high quality. You can tell this is not one of those dull, stale matcha powders. It mixed well and had a very clean, balanced flavor.
As someone who was looking for authentic Japanese matcha without the marketing gimmicks, this felt like a great find. Looking forward to trying it next with just warm water to experience the flavor on its own.
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Reviewed in the United States on May 26, 2026
★★★★★ 5
Smooth like butter - Better than Jade leaf
Flavor Name: Superior Blend, Size: 3.5 Ounce (Pack of 1)
This stuff is smooth. No hint of bitterness in sight. Its so smooth I actually preferred to mix it 50/50 with Jade Leaf in my Matcha lattes. Its just amazing Matcha.. so good I started occasionally drinking it by itself not as a latte... the flavors present in just the tea (no milk, no sugar, etc) is insane. Umami, savory, very minimal mineral/bitter tastes. So if you like mineral tasting matcha, try Dens tea Restaurant Matcha... it has a very strong mineraly taste. This is almost the opposte. One thing to note, this stuff is so smooth and lacks bitterness that I actually have to use 3.5g instead of 3g like I do with Jade leaf... but once you get the strength dialed in it is MINT. I personally dont care about any clumping etc because I whisk my matcha with an electric frother in about 2 oz of water before adding milk or anything else.. even the matcha i had in my fridge which got clumpy from condensation didtn stand a chance... so if you ever have lumpy matcha.. its your fault, not the matchas fault. FYI. I dont mind the Jade leaf at all (for lattes)... in fact I wil gladly use it if its on sale.. but this is objectively the better matcha. If you plan to drink by itself, dont even consider jade leaf.. I am speaking specifically for lattes... Naoki is the only one you want to drink straight. But if you are a latte person.. either one is good quality and a buy again from me.
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Reviewed in the United States on March 20, 2026
★★★★★ 5
Best matcha hands down
Flavor Name: Superior Blend, Size: 1.4 Ounce (Pack of 1), Flavor Name: Superior Blend, Size: 1.4 Ounce (Pack of 1)
My favorite matcha I’ve gotten from Amazon thus far. I love the nutty flavor and it’s so creamy and the quality is 10/10 ! I highly recommend
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 2, 2026
★★★★★ 5
Great quality and value!
Flavor Name: Superior Blend, Size: 3.5 Ounce (Pack of 1), Flavor Name: Superior Blend, Size: 3.5 Ounce (Pack of 1)
After my Japan trip last year, I became a Matcha lover. Bought a few different brands, Naoki is the best and is my favorite. Taste fresh, smooth and love the bright green color. I drink Matcha latte everyday. Also whenever my friends come over, I would make one for them to show off my Matcha making skills. They love it as much as I do.
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Reviewed in the United States on March 26, 2026
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