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Description
Human S100A12 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: 1. 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: 1. 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. 2. 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. 3. 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. 4. 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. 5. 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. 6. 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 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 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 conjugate are sequentially added to microwells pre-coated with S100 Calcium Binding Protein A12 (S100A12) 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 S100 Calcium Binding Protein A12 (S100A12) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human S100 Calcium Binding Protein A12 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | S100 calcium-binding protein A12 (S100A12) is a protein encoded by the S100A12 gene. The protein encoded by this gene is a member of the S100 family and contains two EF-hand calcium-binding groups. Generally speaking, S100A12 has significant anti-infective and antibacterial effects, which are related to its ability to absorb ions. Neutrophils and monocytes/macrophages are the main cellular sources of S100A12, although some epithelial cells and dendritic cells can also secrete it. Certain tissues, such as the spleen and lungs, are rich in these cells and, therefore, in this protein. It is produced intracellularly but is also produced in the extracellular environment, where it primarily exerts its effects. It exists as a homodimer or a hexamer in the extracellular space. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.8 ★★★★★
Based on 9 reviews
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Product Reviews
★★★★★ 5
Reminds us of the value of a college education beyond academic facts and skills.
Format: Kindle
At a time when many students and families are questioning the value of a 4-year college education, Porterfield does a great job of describing the intangibles behind an earned degree. Many people are aware that there is more to a college education than academic facts and skills, but this book presents the case comprehensively. It identifies "soft skills" specifically and gives good examples of how they can be taught and learned in the college environment.
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Reviewed in the United States on May 3, 2026
★★★★★ 5
Loved it!
Format: Kindle
Mia is a ten-year-old Chinese immigrant. She had immigrated to California from China 2 years earlier with her parents, who were hoping for a better life. Unfortunately, the United States in the 1990s was not exactly welcoming to Chinese immigrants, and many struggled. Mia and her parents were no different. After gaining and losing several jobs and facing homelessness again, Mia's parents take a job managing a motel. Unfortunately, the motel owner - who is Taiwanese, not Chinese, as he is quick to point out - is not only overtly racist, he is also exploitive and seems almost happy to point out to Mia's parents that they could be replaced immediately when they question his actions.
But Mia and her parents try to make the best of it. Mia works the front desk when at the motel, quickly learning the ropes and consistently trying to find ways to help the guests and her family. She also becomes friends with the weeklies at the motel - those guests who live at the motel and pay weekly - and they quickly become part of her growing family.
At school, though, Mia struggles. She can't tell the other students - most of whom are white - that she lives in a motel. She struggles in English, though she loves writing, and she also has disagreements with her mother who thinks she should focus on math and forget about writing because "she'll never be able to write as well as the white kids" for whom English is their native language.
Most of the book is a slice-of-life look at running a hotel and Mia's struggle to integrate with the other students at school. Though it may seem to some that too much goes wrong at the motel in too short a period of time, I can say from experience that this depiction is incredibly realistic. While reading it, I was having flashbacks to my own experience managing a motel and running the front desk. The news Mia and her family get in chapter 55, in particular, hit hard on the feels for me, because my husband and I went through that same situation. We didn't use the same solution that Mia and her family did, and I definitely loved the solution they came up with.
Throughout the story were the letters that Mia wrote, each for a different situation. These were an added touch, helping to bring the reader back to the days of the early and mid-90s, before email was ubiquitous and text messaging was still a dream. Her struggle to put word to paper, to make sure she was using the right words and tenses, was an added dimension to the story that I didn't expect but definitely appreciated. The author's note at the end, where she discussed her own experiences was also incredibly moving, and I loved learning that much of the book's scenarios were pulled from her own experiences, while also wanting to give a hug to the child that had to experience that struggle.
I had the opportunity to listen to the audiobook, narrated by Sunny Lu, and I highly recommend it. Lu did a wonderful job with the narration, injecting just the right amount of emotion and tension into the performance. I had to force myself to stop listening so I could go to bed; had I not, I would have listened right through to the end and not getting any sleep. While I would have enjoyed the story, work the next day would not have been fun. Lol.
So, long review short, I definitely recommend this book. For younger readers, I would recommend a parent read with them, as there are some tough situations that they may need/want to talk about. But overall, this is a story of hope for an immigrant family who is struggling to make a better life for themselves.
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Reviewed in the United States on October 18, 2022
★★★★★ 5
A great book for all
Format: Paperback, Format: Paperback
What a great book. I bought it for my 12 year old granddaughter. A story of resilience.
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Reviewed in the United States on March 16, 2026
★★★★★ 5
My favorite middle grade book of 2018 so far
Format: Hardcover, Format: Hardcover
I'll never look at a motel with the same eyes again. Author Kelly Yang brings us Mia Tang, a younger version of herself, who helped her parents manage a motel in southern California in the early 1990s.
Mia and her parents arrive from China with two hundred dollars, unwavering optimism, and the belief that if they work hard enough, they will achieve the American dream: a big house, a dog, and an endless supply of juicy hamburgers. After two years of backbreaking jobs and sketchy living conditions, Mia's parents rush to sign a contract to manage the Calista Motel located five miles from Disneyland.
Mr. Yao, the notoriously cheap and unscrupulous motel owner, informs the Tangs that the contract allows him to make any changes and if they don't like the terms, "Just say the word. There are ten thousand immigrants who would take your job in two seconds" (p. 27).
Mia quickly makes friends with the "weeklies" and assigns herself front desk duty because her parents must clean every room themselves and be available 24/7 to check in guests.
Mia's moxie and sense of justice emerge as she takes on adult responsibilities with a sense of pride and unbridled enthusiasm.
She makes plenty of mistakes which makes us empathize with her struggles and root for her as she tackles the english language, bullies, crime, and embarrassment about her thrift store clothes. (Keep your eyes open for the tale of the designer jeans.)
Chapters are vignettes that are strung along with seamless transitions to make reading a pure pleasure. The author captures Mia's voice so authentically that the reader is instantly drawn into the story. Mia's uses the power of the written word to advocate for herself and others.
Kids will fall in love with Mia and cheer her fierce determination as she navigates her way through poverty and injustice, bringing her family, the weeklies, and everyone else who want to be part of the American dream with her.
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Reviewed in the United States on September 12, 2018
★★★★★ 5
Outstanding!
Format: Hardcover
Hiya Bumbles! I am so excited to share this upper elementary/middle grade book with all of you! This is the story of Mia Tang, a Chinese immigrant, and her family’s quest for a better, “freer” life in America. The Tang family is given the opportunity to manage a motel and a journey of heart breaking and heart filling experiences begin. Ms. Yang artfully brings sass and humor to very serious topics regarding social issues that continue to pervade our communities. This is quite a relevant read right now and I can see how fabulous discussions and lessons will arise from this novel. The main character, Mia, is spunky, bright and spreads love everywhere she goes. She also stands up for herself and others when it counts. I loved getting to know her. It was eye opening to read some of the circumstances that the immigrants in the story have to live through. And although there are heavy social injustices that are prevalent in her life, Mia handles them with grace and just the right amount of sass and compassion. I absolutely would recommend this story and I think it teaches so many valuable lessons to readers! I also think that it can transcend age up through YA and it’s a great read for adults too. It’s actually based on some of the author’s own experiences.
🐝🐝🐝🐝🐝 from me! Loved it!
#frontdesk #kellyyang #scholastic #kidlit #bookstagram #immigrant #chinese #miatang #spunkandsass #booknerd #kellyyangisawesome
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Reviewed in the United States on June 20, 2018