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Description
Rat CX3CR1 ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 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 Chemokine C-X3-C-Motif Receptor 1 (CX3CR1) capture antibody. 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 Chemokine C-X3-C-Motif Receptor 1 (CX3CR1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Chemokine C-X3-C-Motif Receptor 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | CX3C chemokine receptor 1 (CX3CR1), also known as bifurcating protein receptor or G-protein-coupled receptor 13 (GPR13), is a protein encoded by the CX3CR1 gene. As its name suggests, this receptor binds to the chemokine CX3CL1 (also known as neuronal or bifurcating protein). Expression of this receptor appears to be associated with lymphocytes. CX3CR1 is also expressed by monocytes and plays an important role in their survival. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.4 ★★★★★
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Product Reviews
★★★★★ 5
CODE is the book you should get.
Format: Paperback
The best book to enter the field. Grateful that Charles Petzold decided to write it, even happier this was one of the first books I picked up.
Recommend to everyone wanting to become a programmer, or just interested in computers.
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Reviewed in the United States on January 12, 2026
★★★★★ 5
If you only buy one technical computer book, this is it
Format: Paperback
Disclaimer: I owned the first edition for years and read it a dozen times, mainly for pleasure since it’s not only informative but also just enjoyable to read. I’m a professional software architect and have dozens of textbooks on computer science and engineering so I’ve been around the block.
This is the kind of book I would buy all for all my friends (if I had more than a couple) and family members (if they were even slightly into computers, which they aren’t sadly). Petzold does an exceptional job at describing digital logic and guides you through building a conceptual CPU from very first principles.
I think his bottom up approach is the way to go and has helped me understand complex topics in an extremely simple way. When I’m not reading fiction, this is my go to bedtime reading.
The second edition has about 100 more pages than the first and some content has been completely reworked and it’s great.
If you ever had even a passing interest in knowing how computers work at a very fundamental level, get this book!
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Reviewed in the United States on January 31, 2023
★★★★★ 4
First programmer?
Format: Paperback
As an ardent reader of the past 1st edition, I was fairly excited to get the latest version. That is, until I encountered the history chapter of the new edition.
For one thing, what I do NOT understand in this second edition is the newly added description of August Ada Byron (countless of Lovelace). The author claims it was Babbage who was the first programmer to design the engines, not Ada. I am not trying to start a futile argument here about who has more or fewer contributions, etc.
What I am trying to assert here is that it is undisputed that Ada (unless the new evidence arises) left *the very first demonstration* of what this seemingly imaginary machine, which didn't even physically exist, was capable of through her program. Because Babbage designed the engine itself, that doesn't automatically put him in the position of a programmer (despite Babbage being a brilliant engineer/scientist and may have had a simple or detailed program in his mind). However, it was Ada who gave a definite touch to programming concepts that ultimately led to modern-day programming. Ada deserves more recognition than a mere "tutorial writer," and she is certainly entitled to the title she deserves.
Other than that, like the previous edition, this book is a must-read for people who are from related/unrelated fields. I always loved the 1st edition, and I would do too with the 2nd. Still, I think history should always be approached with more care, particularly if matters have potential controversies.
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Reviewed in the United States on January 6, 2023
★★★★★ 5
One of the few books worth your time
Format: Paperback
If you want to learn computers, start here. Learning from a historical perspective removes complexity of trying to start with say GPUs - you learn just like computer engineers did, incrementally.
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Reviewed in the United States on April 27, 2026
★★★★★ 5
An absolutely brilliant book
Format: Paperback
This is an amazing book for the right kind of reader. It's a lot like Euclid's Elements but for computers as it leads the reader through designing a computer from first principles and builds to the point of creating software and adding peripherals. It's an amazingly clear but slightly challenging read. I have given this book as a gift many many times. It's an absolute classic in my opinion.
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Reviewed in the United States on March 19, 2025