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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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4.6 ★★★★★
Based on 6 reviews
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Product Reviews
★★★★★ 5
An Excellent, Eye-Opening Read That Truly Makes You Reflect
Format: Kindle
The 10X rule is an excellent book—engaging, interactive, and surprisingly eye-opening. It doesn’t just present ideas; it invites you to actively reflect on your own behavior, decisions, and patterns. As you move through the chapters, you’re constantly prompted to pause, think, and reassess the way you act and respond in everyday situations.
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Reviewed in the United States on February 8, 2026
★★★★★ 5
Grant Doesn't Hold Back!
Format: Hardcover
What It's All About...
So, the 10 X Rule is a very motivation book about rethinking the way you think about goal setting and the effort it takes to accomplish your goals.
Without giving away the obvious, Grant shares that when we go for a goal like, "I want $100 in the next 30 days to pay my bills." We often fall short of that goal and need to restate it as: "I want $1,000 in the next 30 days to pay my bills." Now, if you do far short (happens often), you will still hit somewhere greater than $100. Therefore, we must always 10 X our goals to hit the goals we desire.
Now, when it comes to effort, because we often underestimate the action it takes to meet our goals... Our new $1,000 goal requires 10 X effort to hit it. So, if you think "To hit $1,000, I need to make 1 sales call each day." you need to 10 X that number and make 10 sales calls to hit your new $1,000 goal. Therefore, we must always 10 X our effort and change our actions to meet out goals.
He also takes times at the end of the book to outline 32 characteristics of successful people that also help in accomplishing these 10 X Goals. Some of those include: Have a "Can Do" attitude, Seek to Solve Problems, Focus on Now, Be Highly Ethical, etc.
At the end of each chapter he asks a set of questions that helps to reinforce the material you just read.
My Take...
Overall, this really juiced me up and got me to rethink the way in which I look at goals and the actions I need to meet them.
When reading such a book, I often get so inspired that I shoot off goals without really appreciating the amount of effort it will take to hit them. I walked away from Grant's book with a firm idea on the big goals I want to hit and a healthier understanding of what it's going to take to get there.
The overall feel of the book was also inspiring because Grant is the type of person that believes in going for goals, even if you don't know exactly how you are going to accomplish them... Just get started. I resonate with that line of thinking.
If you like listening to audio books, Grant does his own reading of the book, which gives a greater sense of emphasis and passion behind the content.
Who should read this...
Entrepreneurs, Business owners, High School and College Students
WARNING: There is some cursing.
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Reviewed in the United States on May 20, 2016
★★★★★ 4
A highly motivating ride built on top of a simple truth
Format: Kindle
Underneath its swashbuckling and action-oriented prose, Grant Cardone's The 10X Rule is a simple book about risk reduction.
In a motivating and positive tone I would describe as "pleasantly aggressive", Cardone cajoles the reader as he sells a relatively simple premise backed up by his own experiences.
Cardone believes that the difference between success and failure lies in executing his 10X Rule, which is:
1) Set goals ten times higher than what you need to achieve, and then
2) Take ten times more actions than you think are necessary to achieve your goals.
That's really it, and this is risk reduction. Understand where you need to aim, then shoot much higher to create a greater margin of error, and then take massive and relentless action to plant an entire field full of seeds, some of which are likely to bear the bounty you are looking for.
It's simply a numbers game.
Cardone writes in a tone that will get you fired up, that will get you ready to take on larger challenges and keep going -- in fact go harder -- in the face of adversity.
The 10X Rule is full of quotes designed to kick you into a higher gear:
"As long as you are alive, you will either live to accomplish your own goals and dreams or be used as a resource to accomplish someone else's."
"One of the greatest turning points in my life occurred when I stopped casually waiting for success and instead started to approach it as a duty, obligation, and responsibility"
In the end, I enjoyed this book and am likely to read it again. The 10X Rule is like an Indiana Jones movie built on an ancient truth. Underneath the testosterone, rah-rah and adventure lies the simple facts: extraordinary results require extraordinary effort. The premise isn't overly cerebral: aim high and go for it. But the pace, voice and storytelling align to motivate, and that's a valuable enough takeaway from a business book.
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Reviewed in the United States on April 11, 2015
★★★★★ 5
Incinerate the competition. (And you have no competition!)
Format: Hardcover
Wow. Just love this book. Grant's ideas are spectacular. I say go with the audio (CD) version because his energy, his voice and his message must be "heard" to make an impact. Ordinarily, the only writers I give five stars to are people like Herman Melville and Joyce Carol Oates. However, IN HIS FIELD Grant is hard to beat. He's articulate and refreshingly straight-on. No b.s. here. Literally, he "sold" me on his ideas (and I am following them, Grant!) I am a writer and artist. All my life I have been told that it is nearly impossible for a cutting-edge visual artist and new novelist to make a good living at what I do. I don't believe that any longer, thanks to Grant's coaching in this book. As a matter of fact, the more unique you are in your field, the more "different" and wild, the more likely you are to be successful because it's likely that you are passionate about what you do. And that's the key to everything: passion. If you believe in your product or service, your talent and your creativity, you can make it because you have what it takes to do anything - even selling what you believe in. A lot of artists think they are "selling out" when they're "selling" themselves. Even Emily Dickinson called publishing "the auction house of the mind." (Or did she say "block"? That would make more sense because the implication is that a published writer is a slave to the publishing industry.) The fact is that most of us cannot afford to split hairs about whether we are selling out or not. We have to make a living to support ourselves, our families, or any of the lives and causes we support (including our selves; it's all about taking responsibility). It's time, guys, to step up to the plate and make it happen. Grant is my coach of choice. I've always gotten a lot from Anthony Robbins, Wayne Dyer and other "success" gurus, but only two at this point take the lead for me: Eckhart Tolle (didn't you say that "NOW" is the only time to move forward, Grant?) and Grant Carbone. I'm sure Eckhart would smile at this. Their styles are completely opposite, but their impact is huge. The only thing I would change for Grant himself is, man, you gotta say your name more on your tapes. I'm almost done with the book and have recommended it to many people. I even bought the book (I got the original from the library) so I could loan it to friends. However, whenever I talked up The 10X Rule (until today) and people asked who wrote it, I couldn't tell them. That's right, Grant, I didn't know your name from listening to your book. Anyway, just thought you should know. Thanks for being in my life. You changed up my game plan tremendously. I.O.U. (Oh, wait, maybe you'd say I just paid with this glowing review!)XXX000
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Reviewed in the United States on February 10, 2013
★★★★★ 5
Great book
Format: Hardcover
Got this book for my husband, who loves to listen and watch Grant Cardone. Great buy and great product. Durable book.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 10, 2026