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Description
Mouse IL-1α 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 them by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with ice-cold PBS and resuspend them 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, or store at -20°C or -80°C, but avoid repeated freeze-thaw cycles. Other biological fluids: Centrifuge at 1000×g for 20 minutes, and remove the supernatant for analysis. Pre-Assay Preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the gradient working solution of the standard: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration is 200 pg/mL). Then dilute to the following concentrations: 200 pg/mL, 100 pg/mL, 50 pg/mL, 25 pg/mL, 12.5 pg/mL, 6.25 pg/mL, 3.125 pg/mL, and 0 pg/mL. Serial Dilution Method: Add 500 μL of universal diluent to each of seven EP tubes. Pipette 500 μL of the 200 pg/mL standard working solution into the first EP tube and mix thoroughly to make a 100 pg/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 utilizes a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotinylated detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with Interleukin 1 Alpha (IL-1α) 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 1 Alpha (IL-1α) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Interleukin 1 Alpha ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interleukin-1α (IL-1α), also known as hematopoietin-1, is a cytokine in the interleukin-1 family, encoded by the IL-1α gene. Generally, interleukin-1 is responsible for producing inflammation and promoting fever and sepsis. IL-1α inhibitors are currently under development to disrupt these processes and treat disease. IL-1α is primarily produced by activated macrophages, as well as neutrophils, epithelial cells, and endothelial cells. It has metabolic, physiological, and hematopoietic activities and plays a central role in regulating immune responses. It binds to the interleukin-1 receptor, which is involved in the activation of tumor necrosis factor-α. The mouse IL-1α cDNA encodes a 270-amino acid pro-IL-1α precursor peptide containing a nuclear localization sequence, an N-acylation site, and three potential N-linked glycosylation sites. Most IL-1α is retained in the cell in the form of precursor proteins, and a portion of unprocessed IL-1α is transported to the cell surface. These membrane-bound unprocessed IL-1α also have certain biological activity and can act on the IL-1 receptors on the surface of adjacent cells in a paracrine manner. | |||||||||||||||||||||||||||||||||
| 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 | 3.12-200 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates, cell lysates, cell culture supernatants and other biological fluids |
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4.0 ★★★★★
Based on 9 reviews
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Product Reviews
★★★★★ 5
Astoundingly Good
Format: Kindle
This is a must have book. It is, beyond a doubt, the best book I have read on military strategy. The author is clear, provides case examples, and more importantly makes this "readable."
I retired with 24 years on active duty and spent 15 more working in PMC's working in austere and conflict environments. THIS book is long overdue.
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Reviewed in the United States on April 6, 2014
★★★★★ 5
Thoughtful and deeply insightful
Format: Kindle
Browse not only goes over the current state of the US military in detail, but also ends with concrete and manageable suggestions to fix the major problems. Really good book.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 19, 2025
★★★★★ 5
Eye Opening, Thought Provoking and Scary
Format: Hardcover
This book will grab your attention, keep you spell bound and scare the heck out of you. The author was the Chief of Staff under Senator McCain for the Senate Armed Services Committee. This book is about new technology in the defense field and our inability to deal with it.
The new technology comes in many forms. There now are missiles that fly 2 or 3 times faster than what is available now. The missiles can reach out many many thousands of miles more, enough to hit America from the other side of the world. Now computers are recently coming out on the market which are smaller and 2 or 3 times faster than previous computers. All of that combines to radically speed up the decision time for war operations. The author calls it the kill chain.
The change doesn't stop there. The tactics used by our competitors has radically changed warfare. The examples the author uses comes from Russia. He reviews their invasion of "Little Green Men" in the Ukraine turned warfare upside down. They infiltrated troops into the land. Then they merged with dissent forces already in the country. Then the war stars, but on a small scale. Before you know it Russia grabbed Crimea and neutralized a huge slice of the Ukraine. That was the first time since WWII where borders changed.
The last part of the book is the most scary. He relies on his experience in Congress. He cites several examples to show where the bureaucracy is incapable of change. The pressures of on going operations, turf wars, political desires to protect home based companies all have immobilized the bureaucracy. He also cites the case of the Army trying to get a new side arm. It took 17 million to test an off the shelf pistol. The case showed how fear of risk has layered on level after level of control and check. Those levels of course adds costs. That was just one weapons program. Can you imagine what the cost is as you expand that out to really big ticket things like carriers. It leads to the Pentagon to continue buying weapons it doesn't need and use tactics which really come out of WWII. As the Pentagon games go on the world's armies change. I think his point about the bureaucracy caught in a never ending loop also might explain other troubles across the globe.
That leads to the scary part. Is the country ready for the future? Will it defend the nation for the future? If it isn't 9/11 might be a match strike in comparison.
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Reviewed in the United States on June 11, 2020
★★★★★ 4
Eyes Wide Open (with a few omissions)
Format: Audiobook
Thought provoking and great insights but with a few material omissions.
As others have noted this is an extremely thought provoking book. Perhaps the most disturbing is the discussion of war gaming a war with China and in most every Chinese initiated war China wins. A close runner-up was the lack of widespread commitment of other senate members to be as fully informed as possible on the military side of military affairs including budgets for specific projects.
It's hard to document the claim that two issues were serious omissions but I think there were.
There are seemingly minor details that are important Robert McNamara worked for Ford not GM. This is important for decisions at Ford by McNamara's accolates took Ford down to one of its smallest market share of the postwar years. McNamara gave Ford the Falcon , his successors brought out the Mustang. His arrogance cost billions and thousands of lives.
McCain recognized the political folly of the initial "leased" Boeing Replacement Tanker Program but that is not discussed. Neither is the continuing debacle of the program, felony convictions/pleas of top Boeing execs and the Pentagon's civilian chief of procurement all associated with the ill-fated tanker program. Declared a near emergency need at the turn of the century, twenty later the tankers can not perform the mission and tens of billions over budget.
To put the Tanker Program debacle in perspective, In July 1962 the US achieved its first orbital space flight and its first Moon landing 7 years later. In contrast the replacement tanker program has been in process Boeing was awarded the contract in 2002 , 19 years later and the tankers are not fully operational. Along the way both Boeing and a top civilian dod official did some hard time on felony corruption convictions/pleas.
The author notes that in the event of an outbreak of war between the US and China the US ships must get far offshore to have even a chance of survival, well beyond the range of existing carrier based aircraft to attack Chinese forces. The lack of tankers, short range attack aircraft and light loads prevents the Navy from going deep inland.
Part of the problem is that the Navy was induced to scrap the long range, extremely deadly F-14B and F-14X and replace them with the slower, shorter range , less carrying capacity F-18s (also made by Boeing) . The Navy had available at the time the F-14X upgrade program which would have converted the F-14 to an even more deadly fighter / bomber and equipped them with a follow-on to the Phoenix missiles, so badly needed to defend the fleet against airborne launched cruise missiles. In addition there were further upgrades in the works to give the Phoenix missiles extremely valuable capabilities. A further indication of the suspicious pattern is that DOD required that all F-14 tooling and parts be destroyed. The claim was made that the F-14s were maintenance hogs. Partly true but largely fixed with the F-14X digital conversion and new engines. While the maintenance hours per flight hour were problematical, when looked at in the big picture they were a rounding error in the 6,000 or so sailors in the Battle Group working 10-15 hour days and the thousands onshore supporting the effort.
Does this matter, well yesterday the Chinese ran a practice attack on a US carrier as about 15 aircraft approached within 250 nautical miles of the carrier. Most certainly within range to launch enough hypersonic cruise missiles to virtually assure the carrier would be taken out of action or sent to the bottom of the ocean. As the author notes today's strategy requires that the carriers flee the area and standoff about 1,000 miles. Faster, much longer range F-14x aircraft with the next generation Phoenix would significantly reduce this threat. They would also do the same against large Russian aircraft carrying many cruise missiles.
The F-35s will help overcome this deficiency but until they are fully operational and our Naval tanker capabilities redeveloped US capabilities are seriously compromised.
The author makes many great observations regarding deficiencies in procurement management, in the Pentagon , Congress and White House.
Examples discussed include the Army's failed attempt to acquire a new pistol. The 500 page request for proposals and flawed competition would be a joke were in not for the fact that the taxpayers precious dollars were wasted in the failed effort.
An illustration of how perverted the situation has become was illustrated today with a note the the US Air Force had issued an RFP for a "modesty curtain" to be installed on our ancient B-52's because there were now female personnel flying missions. This is a need that should be solvable by a few individuals over a bottle of wine who would probably come up with better ideas, reviewed by an engineer on Monday and perhaps fabricated in one of the base shops.
As others have noted it was USAF Col John Boyd who revolutionized the air to air combat, was shunned by top Brass while at the Pentagon and left to his own devices prepared his famous day long lecture on Winning and Loosing Wars that in turn helped rewrite the USMC land battle doctrine. Most all of this work done out of sight of his "leaders" .
The author might have also given credit to leaders like Admiral Tom Connolly who sacrificed his career to save Naval aviation from the terminally flawed F-111B as an example of the character and courage needed in the Pentagon, Congress and the White House today and into the future.
The author's descriptions of the challenges posed by an aggressive and expansive China should be taken to heart by every American. Unless we stop treating military procurement as a Chicago like spoils system and manage both what we buy and what we pay for it we are inviting Chinese military challenges and placing an even greater financial millstone around the necks of American taxpayers and their future generations.
Overall , not perfect but a very important must read
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Reviewed in the United States on January 30, 2021
★★★★★ 5
Add this to every DoD Reading List!
Format: Kindle
There were so many great excerpts throughout the book that my highlights filled 16 pages in MS Word. This is a must read for all Pentagon personnel, those in operational commands, the acquisition community, and defense industry. Chris' insights as McCain's advisor were invaluable to understand the nuances and competing incentives of the DoD, industry, and Congressional perspectives.
"The problem is that America is playing a losing game. Over many decades we have built our military around small numbers of large, expensive, exquisite, heavily manned, and hard to replace platforms that struggle to close the kill chain as one battle network. China, meanwhile, has built large numbers of multi million dollar weapons to find and attack America’s small numbers of exponentially more expensive military platforms."
"It requires a sweeping redesign of the American military: from a military built around small numbers of large, expensive, exquisite, heavily manned, and hard to replace platforms to a military built around large numbers of smaller, lower cost, expendable, and highly autonomous machines."
"New technologies alone will not save us. We need new thinking — an ambitious effort to reimagine the ends, ways, and means of US military power, as well as the role of our allies in this effort — to succeed in a future world where America’s military superiority will likely erode further if China’s military technological development continues."
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Reviewed in the United States on May 9, 2020