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Description
Rat IL-9 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 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 500pg/mL). Then dilute to the following concentrations: 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, 15.625pg/mL, 7.8125pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 500pg/mL standard working solution into the first EP tube and mix thoroughly to make a 250pg/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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| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Interleukin 9 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interleukin-9 (IL-9) is a multipotent cytokine (cell signaling molecule) belonging to the interleukin group. It is produced in varying amounts by various cell types, including mast cells, NKT cells, Th2, Th17, Treg, ILC2, and Th9 cells. Th9 cells are considered the primary CD4+ T cells producing IL-9. This cytokine stimulates cell proliferation and prevents apoptosis. It acts through the interleukin-9 receptor (IL9R), activating various signal transducer and activator (STAT) proteins, namely STAT1, STAT3, and STAT5, thus linking this cytokine to various biological processes. The gene encoding this cytokine has been identified as a candidate gene for asthma. | |||||||||||||||||||||||||||||||||
| 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 | 7.8-500 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.7 ★★★★★
Based on 936 reviews
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Product Reviews
★★★★★ 5
Spectacular Albeit Unknown History of Race Relations
Format: Hardcover
This is a great piece of historiography about something few know about at all --- slavery in New York City in the 18th century. How about a slave "rebellion" in New York City, how about more people burned at the stake than in the Salem witchcraft trials, how about dark byways and highways of old New York, barely transformed from its days as New Amsterdam, dark plots in dank places, shrill frightened tyrants overreacting with bloody retribution, burned ruins of an early African American village in Central Park?
One cannot make up this stuff, it is too real so it must be history at its best.
And written by one of our premier authors of history, a woman who makes our history live in The New Yorker to the acclaim of many, and yet whose best book, this one, is still too little known.
If you appreciate Harry Truman's remark that the only new thing under the Sun is the history you haven't read, then this is one to curl up with and marvel at; a great way to spend a rainy day or a dark night.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 22, 2010
★★★★★ 4
Good, but not great.
Format: Paperback
Kudos to Lepore for delving into an important, little known subject, which she does better than most historians. At times, however, I think she felt the need to put every little piece of information she got into the book. It was way too long. Some good research, but she has done better. Still, worth checking out. I like to think I know American history, but I know nothing about this awful chapter.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 1, 2019
★★★★★ 5
DAMN, this is a great book!
Format: Hardcover
All history books should be this detailed, this readable, this humane. Lepore knows how to write about a horrible, nearly forgotten episode in NYC history. Unlike many historians, she steps away from overt politics or raw emotion. She knows that this subject is too serious to be shouted. It is the rare history book that is packed with facts as well as knowledge.
I felt like Lepore was taking my hand and leading me through the smelly streets of lower Manhattan in 1741, like I could almost see the faces of...what were they, anyway? The victims of a horrible hoax? The demented planners of a plot to burn the city? Or something in between, where thieves can also be the keepers of ancient rites from a distant homeland, where the world is turned upside down?
I could go on and on, but just buy the book!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 20, 2008
★★★★★ 3
New York Burning
Format: Paperback
.
This is an important book that explores in depth what is usually only found in textbooks as a one-sentence summation:
"In 1741 there was a slave uprising in New York City."
Scholars will probably be happier starting with the Appendix and bibliography and then reading the book. The text is disorganized and uneven, and although this is non-fiction, the characters could have been more finely drawn. Peter Zenger's trail keeps popping up in unexpected places, often disconnected from the action the author is working on. Some sections are heavy on primary documents and period writings, others are more poetic.
Yes, I do understand the parallels with the Salem Witch Trials. The Salem Witch Trials get more press today because of Arthur Miller's "Crucible." Color and religion of the participants aside, both events are stories of group think and mass hysteria, fear and anger. There is plenty of room here for a first-class film or play to be written.
Read this book, learn from it. Expect to complain about it.
Kim Burdick
Stanton, DE
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 7, 2014
★★★★★ 5
What You Didn't Know
Format: Paperback
Did you know that if you were a Catholic Priest on the streets of New York in 1747 that you'd be arrested and hung! Great book if you're interested in the times during which our founding Fathers were growing up. It'll give you a different concept on how slavery was different in NYC as opposed to in the South, and how many of the streets in NYC got there names from English magistrates. If you like history, especially of NYC, you'll love this book.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 24, 2015
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