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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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4.9 ★★★★★
Based on 352 reviews
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Product Reviews
★★★★★ 5
Great card to have highly recommend
Like using the card for everything. 5 to 7% cashback credit is awesome.
No interest is even better.
I put almost all my purchases and bills on my credit accounts or credit cards and pay them off monthly.
Now that I'm a senior citizen I don't like to go to 20 different stores to find the best prices.
The card made my life less busy and I don't have to worry about paying for the many, many things I need right away. Even though I pay off all my accounts every month.
If I need a big ticket item, like a new flat-screen television I can get it and not worry about the cost by paying over time if I need to.
Having another credit account has also improved my credit score by the hundreds. I was in the 500s for decades until I started the secured card on Amazon. Now I am nearly at an excellent score in the high 700s and don't need secured credit cards.
I can use my account at Whole Foods and have my phone scanned for payment. It's just so easy to use
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 6, 2026
★★★★★ 3
Credit limit drop without cause, Synchrony?
Hey Synchrony, I've about had it with your services, I have 2 cards I use through you through Walgreens and Amazon. You lowered My credit limit on both cards causing me to go over on one because of stupid interest when I had 500 left and I had quit using the damn thing, and my Amazon store card I just Made a 200 Payment on! Lowering my limit and leaving me blind, You should call your customers leave a voicemail or text for heck sake ill read them, I don't read my emails usually for 2 or three weeks, well you did it again, but this time to card I just paid 200$ on too!, Your company sucks, I hate your practices, and I want you to know that. I also want Amazon to know that!! I spend a Boatload of money through amazon, as does my entire family, all ya gotta do is look!! Well I have never felt more unappreciated, insulted, like I stole something or like dirt ever ever in my adult life like you made me feel. Like i'm doing something wrong. Well guess what Synchrony Bank? I'm not doing anything wrong, at all!! I've never been late or missed a payment with my Amazon card or any card I have with you, Synchrony. Your Faceless Corp in its infinite wisdom, decided it would be a great idea to screw over an actual good customer just because some stupid algorithm reported my credit limit was too high and I was a risk, and lowered my limit from where it had been for the last 3 years and then sent a Quiet email about a letter that you put on my online account, please read it, just like that, no Urgent header nothing not that. I don't even read my email that often. I DO read texts or my regular mail, or listen to my phone messages!! I meant what I said NEVER LATE not even once, and NEVER MISSED a payment NOT once!!! The ONLY time I ever had to deal with any kind of problem was the one that lowering my limit on my Mastercard were the cause of, Synchrony!!! when you dropped my credit limit without me knowing and I went over my new dang limit because interest charges were coming in!!! I new where I was and had it handled and would have been able to ride the storm but no you had to mess with my account. Oh and I dont give a Crap if you quote me "we have the right to raise or lower you limit whenever we want", It's a BS excuse to screw over your customers, I know about the terms, however in all of my 54 years, you are the only credit card company that has ever lowered a limit one a CC I had EVER! and I've dealt with some Super sketchy Dirtbag Why did I ever agree to this card, charge me for everything Companies. Mostly when I was young but not even they cut my limit, without me missing payments or being late .....Now 30 years later much older and I thought wiser better CC much better 2500$ limit I've had for years and expect that limit to stay in place or like my Prime Store Card 2000$ limit to stay in place!! not dropped by 500$, for hells sake!!!! Well today I get the old hey your payment option for your try before you buy order needs addressed, and why do you think that was? Hmmm? Well, Ill tell you!! Because you Dropped my limit, by 500$ Now if Amazon dropped it then Ill chew them out too, but I don't think it was them, cuz you did this on my Walgreens card too!!! I just made a 200 payment on my Amazon card as well jerks! Not one of the several companies ive had previous Cards with NOT Cap one Cred one, citi, Community bank, american express, discover you name it, and Ive probably had it at one time or another dozens upon dozens over my adult life at this point anyway. Well, not even the Sleaziest Dirtyiest Gutter Scum CC company with fees for just looking at your card has ever lowered my credit limit not in 34 years Especially when I wasn't late or missing payments ever, always been on time and never missed a payment not 1 time, Except SYNCHRONY, and I think you suck for that!! The only reason I gave 3 stars is Amazon doesn't deserve a 1 star review because of your company! I love amazon, I love their products, their deals their delivery Amazon Prime, im ride or die with Amazon, my kids do all their shopping on amazon they even use it for groceries and stuff occasionally, but I really, really don't like how you treat customers Synchrony, especially customers, that pay on time and even early, oh and on auto pay for Christ sake!!! Now I know why some young dumb kids and people say screw it and stop paying CC bills, cuz the Corporations that pass em out like candy to kids, I might add, are SCUM bags, thats why!! It's all about the GREEN, more, and more and more money its never enough!! Well, when you dropped my credit limit without me knowing and I went over the limit of a different card I had with you, it was because of interest fees coming in. I expected and knew about these fees, however, I also thought I had 500 more than I had available on my card!! What a great day that was, let me tell ya!! I didn't even miss a payment I just didn't make a big enough one to cover the interest that came in, and I also never just pay the minimum payment with these cards either!! Your practices are shady, and the crap you pull against loyal customers that pay on time and even on Autopay are so Scummy!! You know when you dropped my credit limit without me knowing and I went over my limit because of interest, I even reached out to my current lawyer about Bankruptcy choices and stuff and if I should deal with you that way, but I got over bein mad! You know if I was late all the time and a crappy customer missing payments I wouldn't blame you for anything and at least Id know my cards were lowered and probably locked ...but I didn't want to do that, I wanted to try and slide right back into auto pay and get back on track and to deal with a recent Identity theft issue im having and not stop paying my bills OH..yeah, you smellin what i'm sellin? Well because of some terrible thing I must have done in a Past life, I guess. Several months ago now when I was dealing with the death of my mom very suddenly. I also noticed my credit score started dropping and I had no idea why, well turns out i'm MR popular. I had all sorts of crap in my name most of it local where I live and surrounding locally. 100 points in like 5 months!! I digress, Whatever! I don't care anymore, cancel my cards if you want, I'm done! This world is goin to hell in a hand basket, Civil War is almost immanent, a Faceless mob can destroy peoples lives just by canceling them on the internet at a whim. When its true I agree with the mob but when there isn't any proof and someone's life is upturned and then it turns out to be a total lie or they said something 12 years ago and its dug up? Good God, I probably said some haynous stuff when I was 25, but I can assure you when I was 37 I was not that person anymore, my views had changed on many things,,,,that stuff is crazy, and so am I for thinking I can do a damn thing,,,, my mom died 5 months ago, I was just recently laid off from my job I had for 17 years last month, and now you drop my limit again when i've been on time and auto paying and almost screw up my Amazon account!!so screw it!! Eat It Synchrony Bank,Eat It! Sorry about 3 Stars Amazon.
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Reviewed in the United States on March 24, 2024
★★★★★ 5
Game changer for clogged pores
Style: New formula!
Game changer for clogged pores and very sensitive, acne prone skin. Saved my rosacea too! Over time, avene products have literally transformed my skin and this was a vital contributor to that.
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Reviewed in the United States on May 6, 2026
★★★★★ 5
Great fragrance & texture!
Style: New formula!
I bought this because I loved the spa-like fragrance of the Avene micellar water. This also has a wonderful fragrance and texture, and love the way my skin looks and feels after Inuse it.
Thank you for being environmentally conscious and vegan as well.
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Reviewed in the United States on August 13, 2025
★★★★★ 5
Super smooth and gentle cleanser
Style: New formula!
I love Avène’s products. This face wash is super gentle and hydrating and I love to use it in the morning to wash my face or at the end of the day when my face feels super dry (otherwise I use the Avène acne cleanser) - it does not leave my skin feel stripped or dehydrated.
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Reviewed in the United States on June 11, 2024
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