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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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4.8 ★★★★★
Based on 2161 reviews
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Product Reviews
★★★★★ 4
Enchanting
Format: Kindle
"Queen of Roses" by Briar Boleyn is a delightful and refreshing reimagining of the classic tale of King Arthur, with a captivating twist that places the spotlight on Morgan, a character who has often been overshadowed in traditional retellings. Boleyn's creative decision to shift the narrative perspective to Morgan breathes new life into the story, offering readers an intriguing and compelling look at the Arthurian world from an entirely different angle.
One of the most commendable aspects of this book is its incorporation of Fae elements, which adds an enchanting layer of magic and mystery to the already familiar Arthurian setting. Boleyn skillfully weaves the world of the Fae into the narrative, creating a captivating backdrop against which the events of the story unfold. This addition not only adds depth to the world-building but also provides ample opportunities for twists and turns that keep readers thoroughly engrossed.
However, while the book boasts numerous strengths, it does have one noticeable flaw: the characterization of Morgan. While it is reasonable to create a flawed and complex protagonist, it appears that at times, Morgan's character becomes overly difficult and hard to relate to. Her persistently negative perception of one of the main male characters, who is a potential love interest, despite his efforts to support and assist her, may come across as somewhat irrational and could test the patience of some readers. Striking a balance between a strong, independent character and one who can recognize genuine support and affection could have enhanced the overall reader experience.
Nonetheless, the allure of "Queen of Roses" lies in its innovative approach to the Arthurian legend and its skillful blending of fantasy elements into a familiar narrative. Boleyn's evocative prose draws readers into a world where magic, destiny, and fate entwine, leaving us eager to uncover the mysteries that unfold within the pages.
I received a free copy of this book via Booksprout and am voluntarily leaving a review.
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Reviewed in the United States on July 28, 2023
★★★★★ 5
An action-packed dark romantasy
Format: Kindle
I loved this book! Queen of Roses is an Arthurian-inspired dark romantasy that is the first book in the Blood of Fae series. The story follows Morgan, the princess of Camelot who is rumored to be part fae. Fueled by prejudiced hatred and a mistrust of fae blood, Morgan’s abusive father strips her of her birthright and hands it to her half-brother, Arthur. Instead of becoming queen, Morgan is commanded to join the temple of the goddesses when she comes of age.
However, Arthur turns into a psychopathic, power-hungry, fae-hating king as he ages. He develops malevolent plans and commands Morgan to find an ancient weapon with legendary power. Although Morgan is wary of Arthur’s intentions, she embraces the opportunity to go on a journey and potentially change her fate.
The story picks up from there and we follow Morgan on her quest to find the ancient relic. It’s full of high stakes adventure, mystery, tension, banter, forced proximity, hidden magic, self discovery, and betrayal. This first installment of the series intricately develops the world building and character development. There’s little romance in this book, but it is evident that it is a slow burn that will continue to develop throughout the remainder of the series.
Overall, I loved the world building, the epic fantasy, Morgan’s journey of self discovery, and all of the twists and turns that set the stage for the future installments. I can’t wait to see what happens next!
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Reviewed in the United States on April 7, 2024
★★★★★ 4
Arthurian Fae Quest…say less.
Format: Kindle
A fae centered Arthurian tale unlike any I’ve read so far. The author did a great job at descriptive world building, with scenes easily playing out in my minds eye. There was plenty of action, suspense, and even a touch of horror. An enemies to lovers, slow burn romance, a quest, with plot twist and turns aplenty. There was a love triangle, which I’m not usually a fan of but, it played out well in this story line. The FMC, Morgan Pendragon, was so blatantly naïve, yet I typically expect as much in a ‘book one’ of a series, especially one that features a fairly sheltered princess. I was happy to read that in spite of this, she still showed a strong sense of morals, fire, and spine. Now our MMC? Kairos Draven, aka Void’s Edge. Oh, how I’m a sucker for a smoking’ hot grumpy warrior alpha with a witty mouth, and a strong sense of “touch her and die” attitude, so you know who held all my cards. That ending? Just made me swoon all the harder. Now add a battlecat that rivals the size of a horse…and well Ms. Briar Boleyn you have well and truly stolen my heart. I’m excited to see where the story goes from here, and follow along to see more of the characters growth. I went into this story fairly blind, and I think I enjoyed it all the more because of it.
Once the story got going, it had me in an absolute chokehold and it was difficult to put down.
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Reviewed in the United States on May 12, 2024
★★★★★ 3
Not a bad start
Format: Kindle
3 stars
Thank you Netgalley and Briar Boleyn for the ARC!
A camelot/king Arthur retelling with fae. I was hooked by the idea of this book immediately and was eager to jump into this world.
• slow burn
• enemies to lovers
• who did this to you
Morgan Pendragon watched her mother die by her father's hand when she was just eight years old, hiding under the bed. Morgan is believed to have the tainted blood of the fae in her veins and is cast aside so that her fathers illegitimate son, Arthur, can become the king. She's seen his cruel treatment of the fae firsthand, so when he sends her on a journey to find a fae weapon she seizes the opportunity to do more with her life. Along the way, she finds more than she could have imagined.
I don't know a whole lot about King Arthur and Camelot but I had a lot of fun with this story! The plot has some similar tropes to popular romantasy books (From blood and ash) but there's enough originality here that it doesn't feel like I'm reading a copy. I liked how the fae were different in appearance than what is typical in most fantasy books I've read. In this book they have blue hair, violet skin and a wide range of other characteristics. I thought that the world building was easy to follow and I could easily immerse myself into this world. After reading the blurb I kept wondering when she was going to go on the journey to find Excalibur and it doesn't happen until around the 45% mark.
The story is a bit slow at times but starts to pick up once they begin their journey to find Excalibur. The John Wick style Inn was a fun concept that I enjoyed reading about.
There are a lot of similarities to this and FBAA and I would have liked to have it be a little more different, but I'm hoping book two will have the story turn into something of its own. Overall I enjoyed reading this story and I'm looking forward to reading book two especially after that ending.
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Reviewed in the United States on May 27, 2023
★★★★★ 5
Astonishing, Fun, Entertaining, Fantastic
Format: Kindle
I consider The Martian my favorite fictional novel of the last 15-20 years. The movie was incredible in that they actually followed the book closer than 99% of other films based on books. It remains my favorite movie of the last 15 years or so as well. I don't know anyone (personally) that loves either of them as much as I do.
With that said, I was REALLY looking forward to Artemis. It was good...but, it was certainly not in the same caliber as The Martian was (at least not for me). I enjoyed it a lot, however and appreciated how author Andy Weir chose to go in a completely different direction and not just rehash another similar story, which I am certain would have been great as well.
As a result, I was cautious regarding Project Hail Mary. It sounded a little too close to The Martian, but yet, also different in that the circumstances simply could not be more opposite and the stakes so much higher. I'm trying to figure out the best way to summarize without giving too much away from this utterly compelling novel. As I read several reviews, I noticed a recurring theme: SCIENCE. Lots and LOTS of science. Holy cow, they were right. Many years ago I read Apollo 13 and Jim Lovell and his co-writer, try as they might, simply could not dumb down Orbital Mechanics anywhere near enough for me to have even a minor clue as to what they were attempting to say...I just skipped 90% of it and hoped that the sentences written afterwards, would help to make sense of what I had just skimmed over. I'm a lot of things, but a math wizard is definitely not one of them. Michael Crichton (Jurassic Park) had an amazing talent for dumbing-down the science of what he was trying to explain in ways that genuinely made sense (most of the time). Not everyone has this talent, and I would say Andy Weir falls squarely in between. He's certainly better than Jim Lovell, but not quite as good as Crichton. But then again, outside of a science textbook, I haven't really read anything with quite as MUCH science as Project Hail Mary. So maybe he's just as good, but he just puts more science into his books than Crichton, maybe that's it...? Either way, be prepared for a lot of astonishingly interesting science within the pages of this novel...and I DO mean a LOT. I don't say this to make you wary or steer you away...on the contrary, Andy Weir has a special talent for making hard science truly entertaining.
The book opens with an absolutely amazing and frightening premise: an astronaut awakes from an induced coma to find the only other two people on board have died at some point along their journey...but it gets worse. He has no idea who he is, or why he's on the ship, and oh yeah, they look to be a long way from home. A really, REALLY long way from home. In fact, the sun he sees isn't actually OUR sun at all. He's managed to leave our solar system entirely. And he has no idea why.
((Minor Spoilers)) The book goes through some clever flash-backs, which set the stage for why the mission happens, and slowly, carefully explains how they managed to get so far away from earth in such a short amount of time. Basically, earth's sun seems to be dying. At the rate of decay, we have maybe 19 years left before the gradual cooling has catastrophic consequences resulting in the death of billions (best guess). Why the sun is dimming is quite the conundrum in the first place. Turns out it really isn't dying, it's being killed by an outside source...which turns out to be easily the greatest find in history. It's alien life, and they are using the sun for food, essentially. It's alien life, but not intelligent life. But still, wow! ALIENS, right???
After this monumental discovery, and some tremendous research done by the most improbable scientist, the investigation into what is happening and why and what to do about it expands exponentially to other nations in order to pool all the resources possible to hopefully save the sun, and by extension, the human race as well. They learn. A LOT. A plan is put together, and with the help of the newly discovered microscopic alien life, which can also double as a power source (along with a few other nifty surprises), they begin to create one last, Hail Mary that could very well be the last chance we might have to save earth. It's audacious. It's dangerous, and it is absolutely critical that it succeed.
As our astronaut's memory slowly unravels, so does his identity: Ryland Grace. He's a teacher on earth. Just a science teacher. Not even a college professor. He's amazingly smart, though. But he's no astronaut...and certainly not one who would volunteer to go on a one-way mission to another solar system to "try" and save humanity. Yet here he is. Alone. light years from earth, trying to solve the biggest riddle in all of human history. Ryland accepts his situation, such as it is, with relative indifference (for the most part). It doesn't matter HOW he got here. He's here now and he may as well use that time to be as productive as possible, right? Along the way, he unravels even more information regarding the microscopic alien life which is slowly dimming our sun during some additional flashbacks. The aliens, dubbed, "Astrophage" are quite the galactic plague as it turns out. Stars all over the galaxy are also losing their light, all due to the little buggers. All that is, except one particular star named, Tau Ceti. Now why would that one star be unaffected by Astrophage, when every single star around it has been affected to some degree. The plan is to go there and figure it out and send the information back, hopefully in time to save the sun before the damage to earth is beyond repair.
There is an incredible amount of stuff going on. The story switches from Tau Ceti to flashbacks of how the whole mission was planned and implemented (which is VERY entertaining, especially Director Stratt, who may actually be my favorite character in the entire novel). Weir is becoming quite adept at building tension, and abruptly switching the story from Tau Ceti back to earth and building more of the backstory then switching back to Tau Ceti. Keeping it all in check and most importantly, interesting all while mixing in a healthy dose of science, which I am to understand is pretty much all genuine, is quite the juggling act. I have long known science can be astronomically entertaining (see what I did there?) when done right...but unfortunately very few people in a position to teach science actually know the best way to create that interest in others. I can say without reservation, Andy Weir definitely knows how to do it...at least in written form.
There is so much I want to say more regarding this truly phenomenal story, but I simply cannot without ruining a lot of the fun and surprises revealed along the way...and it is killing me to keep it locked in. Though I labeled a spoiler warning earlier, I don't think it gave away any more than what the author himself has revealed in interviews he has done regarding the book, and what you can glean from reading the summary here and just a couple other reviews. Tying all of that science together is truly astonishing to me. The creativity to put it into a novel that is remarkably exciting to read is nothing more than incredible talent. Kudo's to Andy Weir for not just hitting a home run, Project Hail Mary is a Grand Slam all the way. I truly did not want this story to end. By the way, I enjoyed the ending quite a bit. I don't know if everyone will. But it was fine for me. I think the ending screams "sequel" at some point too. A lot was left open-ended (IMO) and I wouldn't mind reading a follow-up to this. It doesn't HAVE to happen, but there are a lot of ways where the story could go if Andy chose to do it. Just sayin'.
Just run out and buy this book.
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Reviewed in the United States on May 10, 2021