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Description
Rat IL17A 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 1000pg/mL). Then dilute to the following concentrations: 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, 15.625pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 1000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 500pg/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 utilizes 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 Interleukin 17 A (IL17A) 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 17 A (IL17A) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Interleukin 17 A ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | IL-17A is a protein encoded by the IL17A gene. In rodents, IL-17A was once called CTLA8 because of its similarity to a viral gene (O40633). It is a proinflammatory cytokine produced by activated T cells. This cytokine regulates the activity of NF-kappaB and mitogen-activated protein kinases. It stimulates the expression of IL-6 and cyclooxygenase-2 (PTGS2/COX-2) and enhances nitric oxide (NO) production. Elevated levels of IL-17A are associated with several chronic inflammatory diseases, including rheumatoid arthritis, psoriasis, and multiple sclerosis. | |||||||||||||||||||||||||||||||||
| 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 | 15.6-1000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.7 ★★★★★
Based on 23 reviews
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Product Reviews
★★★★★ 5
Excellent book on ML
Format: Paperback
This is a great book on machine learning. Topics covered are extensive - from beginner level to advanced topics including math behind different algorithms. However, not "all" algorithms are covered. Please go through the table of contents.
The first part - 11 chapters - covers machine learning concepts and second part covers advanced topics with Pytorch. There are lots of excellent code and they work!!
The quality of the book I received is excellent. I have gone through all 742 pages, and it has held up very well!!
I used Jupyter notebook to run all examples. I created a new notebook and copied and pasted the code and ran them. This approach worked very well for me. At the same time, I could experiment with my take on the code snippets and definitely added to my knowledge.
Only issue I have is on the second part of the book discussing PyTorch: (1) Some packages are a bit older version: e.g., transformer 4.9.1 whereas current version is 4.48+. It took some tweaking/recoding to get the examples working. (2) There is not much discussion on why certain architecture was chosen - e.g., number of layers, is there a rule of thumb on how to improve performance by changing these parameters? Even with CUDA the code run for a long time. Therefore, experimenting with different values of parameters become too time consuming. (3) On the same note, if I can achieve test accuracy of 90%+ using logistic regression and almost the same (perhaps one or two percent better with PyTorch with IMDB movie review dataset and that two much faster why should I use PyTorch for this dataset? Obviously, PyTorch is for certain types of problems. Discussions can be included by not adding to the exhaustive (and apt) contents.
Personally I was disappointed by lack of any example on time series.
Must have for ML practitioner as a reference and guide.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 20, 2024
★★★★★ 5
Excellent Textbook for Hands-On Learning of ML
Format: Kindle
This textbook is for the serious life-long learners of machine learning. There are at least two ways to ‘consume’ this book.
For the expert in ML, this is a textbook to study as a clear comprehensive ML overview and then to dive into sections of interest or ignorance. The concepts are grounded in code examples and are well cited (with links) to sources. Further, this textbook is appropriate if you are TensorFlow-centric and want to broaden into cutting-edge ML models/tools coded in PyTorch.
For a new learner to ML, this is a textbook to DO (not just READ) with hands-on and brain-engaged. If you realize that ML is a key life-long skill for your career, consider this textbook as part of a daily learning habit (10-30 min).
From personal experience, my advice to the new learner is as follows… First, clone the GitHub repository, setup your Python environment, and study the textbook, while working through the notebooks. Go on tangents and break the code. Do this methodically as part of your daily learning habit, but do not hesitate to jump ahead several chapters to prepare for tomorrow’s meeting. There is enough excellent material here for a full year of ML adventures.
I did a similar strategy with Raschka’s first textbook. About four years ago, I had finished Andrew Ng’s Deep Learning Specialization as a student in his first cohort. I knew the concepts well but could not do the actual application coding. I was surprised how my Python coding improved by following Raschka’s clean and elegant style. And Raschka’s code examples were meaty enough to be springboards into working applications.
Several textbook editions later, what is different about this new edition?
First, it moves you through scikit-Learn (a firm foundation) to PyTorch, instead of TensorFlow. PyTorch is a better stepping-stone, both conceptually and practically. With PyTorch, you will go further with less energy, while being able to convert your efforts into TensorFlow as needed. In addition, most of the cutting-edge ML/AI/DL research is in PyTorch. It is nice to read a recent arXiv paper, clone their repository, click on the Colab tutorial, and replicate their experiments, along with picking up a ton of new coding tricks & tips. I am excited to work through these PyTorch sections to hone my skills.
Second, there is a clear recognition of model tracking and tuning practices. This is often a gap in other ML textbooks and courses. Once you progress beyond the simple demo examples in a lecture, you realize that the real work is experiments, more experiments, and still more experiments, so that you must understand what the model architecture and hyperparameters are doing to your dataset. There is good coverage of scikit-Learn pipeline, grid search, model performance, and the like.
Third, ML/AI/DL practice is rapidly evolving. Every week new ML packages/services become available that could save much grief on your current project. What is refreshing about Raschka’s textbook series is that he constantly adding cutting-edge topics because he likes to stay current and to help us stay current. Hence, this edition contains recent ML treats as: transformers, self-supervised learning, autoencoders-to-GAN, graph neural networks, DBSCAN, t-SNE (with brief mention of UMAP), and PyTorch-Lightning.
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Reviewed in the United States on February 26, 2022
★★★★★ 4
Just learning it
Format: Paperback
Nice learning book just have to finish it
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 10, 2025
★★★★★ 5
Very useful book
Format: Paperback
I use it for the machine learning class I teach.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 3, 2026
★★★★★ 5
Cover many areas in detail and recommendations for more to read for what's outside
Format: Paperback
Good book!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 4, 2026
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