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Description
Human FceRII 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. 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/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, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a capture antibody against Receptor II for the Fc Region of Immunoglobulin E (FceRII). After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Receptor II for the Fc Region of Immunoglobulin E (FceRII) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Receptor II For The Fc Region Of Immunoglobulin E ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | IgE-Fc fragment receptor II (FceRII), also known as CD23, is a "low-affinity" receptor for IgE, an antibody isotype involved in allergy and parasitic resistance, and is important in regulating IgE levels. Unlike many antibody receptors, CD23 is a C-type lectin. It is present on mature B cells, activated macrophages, eosinophils, follicular dendritic cells, and platelets. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.7 ★★★★★
Based on 9 reviews
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Product Reviews
★★★★★ 5
Very Good Godot Book
Format: Paperback
Great book overall!
Best suited for beginners as it literally begins by conceptually introducing variables, operators, etc. Still, it's very Godot. And nicely up-to-date as well, focusing on Godot 4.x and GDScript 2. Note though that the book only covers 2D, which is a shame. Hopefuly, the author is hard at work on a text thast starts where this one ends and moves on to explore 3D in Godot.
The illustrations are little muddy, but fortunately, Packt offers free downloads of (much crisper) PDFs with proof of purchase.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 25, 2024
★★★★★ 5
Book Review: "Learning GDScript with Godot 4
Format: Kindle, Format: Kindle
Learning GDScript with Godot 4" is an invaluable resource for both beginners and those looking to solidify their understanding of Godot's scripting language. This comprehensive guide takes you from the very basics to creating portfolio-worthy projects, making it a perfect starting point for aspiring game developers.
One of the book's standout features is its in-depth approach. It covers the fundamentals thoroughly, ensuring that readers build a strong foundation in GDScript. The inclusion of numerous code examples is particularly beneficial, providing practical insights into how to implement various features and mechanics within Godot 4.
Visual learners will appreciate the thoughtfully placed pictures throughout the book. While not overly abundant, these visuals effectively complement the text, helping to clarify complex concepts and enhance understanding.
Despite Godot's increasing support for C#, the focus on GDScript in this book is well-justified. GDScript is tightly integrated with Godot, making it an excellent choice for rapid development and learning. The book's code-centric approach ensures that readers gain hands-on experience, which is crucial for mastering any programming language.
Overall, "Learning GDScript with Godot 4" is an excellent resource for anyone looking to learn or deepen their knowledge of GDScript and Godot 4. Its thorough coverage, practical examples, and supportive visuals make it a must-have for your game development library. Whether you're a complete beginner or looking to fill in gaps in your knowledge, this book will guide you every step of the way.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 28, 2024
★★★★★ 5
An Excellent Introduction to Game Programming using GDScript/Godot
Format: Paperback
You’ll get up and going quickly with an overview of Godot and then jump straight into GDScript by learning about variables, control flow, collections, loops, object-oriented programming, and concepts like DRY, KISS, etc. Then the fun begins by building game mechanics while learning specifics of Godot such as the node/scene system, reusing scenes, cameras, collision detection, creating user interfaces, and more.
It also covers local multiplayer where you'll learn the basics of computer networking followed by learning about built-in multiplayer nodes provided by Godot. The final chapters cover some advanced programming patterns like event bus, object pooling, and state machines. By the end, you’ll have a fully playable single/local multiplayer game in the style of Vampire Survivors!
I thoroughly enjoyed my time with this book and highly recommend it for anyone who wants to get started on their journey to learn programming and even for experienced game programmers who want to learn Godot and GDScript.
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Reviewed in the United States on May 19, 2024
★★★★★ 4
Fine so far.
Format: Paperback
I am looking for some of the code from exercises, but can't find it in the project files, and they say don't copy it from the ebook...?
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 17, 2024
★★★★★ 5
Very comprehensive and approachable
Format: Kindle
This e-book covers everything starting from basic general programming concepts and Godot development, going into (relatively) advanced game development programming patterns that are used throughout the industry. It explains not only what the best practices are but why they are used in a simple and understandable way, and provides points of reflection and knowledge testing with quiz sections.
I highly recommend this book for anyone wanting to learn game development as a hobby, anyone interested in learning Godot, and even for anyone trying to learn game programming in general and wanted a simple place to start.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 11, 2024
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