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Description
Human IL8Ra 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, biotinylated detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with Interleukin 8 Receptor Alpha (IL8Ra) 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 Interleukin 8 Receptor Alpha (IL8Ra) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Interleukin 8 Receptor Alpha ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interleukin-8 receptor alpha (IL8Ra) is a chemokine receptor. The protein encoded by this gene is a member of the G-protein-coupled receptor family. This protein is a receptor for interleukin-8 (IL8). It binds to IL8 with high affinity and transmits signals through a G-protein-activated second messenger system. This protein inhibits the migration of embryonic oligodendrocyte precursors in the developing spinal cord. This gene, along with IL8RB, another gene encoding a high-affinity IL8 receptor, and IL8RBP (a pseudogene of IL8RB), form a gene cluster in a region mapped to chromosome 2q33-q36. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.4 ★★★★★
Based on 11 reviews
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Product Reviews
★★★★★ 5
Listened to 4 times and still learning things. Bible based approach.
Format: Audiobook
Listened to 4 times and still learning things. Good approach with lots of Biblical references and basis.
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Reviewed in the United States on October 18, 2025
★★★★★ 5
What is anger ?
Format: Paperback
Helpful descriptions and solutions about anger.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 11, 2025
★★★★★ 5
Read this book slowly, prayerfully and watch as God works in and through your life.
Format: Paperback
How People Change is written by Timothy S. Lane and Paul David Tripp. Both men are experienced Pastors, counselors and Professors. Every year tons of books come on the market that seeks to help readers grow in understanding their problems and overcoming their difficulties. On the back cover of the book Dr. Mark Dever offers this endorsement of How People Change: This book is applied theology. It is about heat, the cross, and fruit. It's about present grace. In sixteen short and well-illustrated chapters, the wonderful prospect of change for the good is held out for the reader. We are called to consider our circumstances and our responses to them, and beneath that to examine our hearts' desires and to afresh to Christ's Cross.
This book will challenge you and confront you in ways that will make you uncomfortable. This is not a book of "feel good self-help" where you will learn six to ten steps to be a better person. This book begins with proclaiming the bad news that man is a sinner in need of a Savior and then proceeds to explain the remedy to sin which is found in Christ alone.
Perhaps you've read a self-help book before, or you've listened to other self-help guru's on TV, and found their advice to be wanting. Drs. Lane and Tripp in How People Change lead their readers through what biblical change is by first helping the reader see their sin so they can behold the Savior who longs to change you through and through. In this book, you will learn to see how God is at work to make you the person you were created to be. In this book, you will be confronted to examine your life in light of the Bible which will be painful at times.
Most importantly this book will help you grow in the Gospel. This book will help you grow through the trials we all experience by not relying on yourself but on the Gospel. This book will help you grow in effectiveness in ministry, because the message of this book is aimed to help you grow in personal holiness unto the Lord. Perhaps you have a struggling marriage- this book will help address heart issues rather than surface issues that are affecting your relationship with God and others. This book will address, confront and challenge your pride, your self-reliance, and your self-righteousness. Yet, this book never misses a beat when it discusses the bad news but also presses the reader to go lay down their burdens, cares, wants, needs, anxieties and problems at the Cross of Christ.
Dr. Dever is correct in his description of this book- it will challenge you to consider your circumstances, your responses to them, and beneath that to examine your hearts desires and turn afresh to the Cross. Take up and read this book! Read this book slowly, and digest its message. Read this book prayerfully and discuss it with others.
I heartily recommend this book for new and mature Christians. The authors explain the Gospel in a way that is thoroughly biblical and yet practical. The strength of this book is that it is thoroughly Gospel-centered.
As I read this book, I was challenged and confronted by areas in my own life that needed to change. As I continued reading the book, and finally finished the book, I saw more clearly how God was at work in my life. The authors achieve their Gospel-goals wonderfully in this book. I can only pray (as I'm sure the authors do and many others) that many more believers take up this book, and not only read it but heed its message.
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Reviewed in the United States on January 19, 2011
★★★★★ 5
Highly recommended!
“How people Change” is the second best book I have read this year (the first being “The Gospel according to Jesus” by John MacArthur). There is indeed a “gospel gap” (as the authors call it) among Bible-believing Christians today. We believe the gospel for our past forgiveness and future hope, but often forget that the gospel applies to present everyday living as well. They focus our problem on heart idolatry - we have replaced Christ with something else! They put it this way – “The heart is the steering wheel of every human being. Everything we do is shaped and controlled by what our hearts desire. That is why the Bible is very clear that God wants our hearts. Only when God has your heart does he have you. As much as we are affected by our broken world and the sins of others against us, our greatest problem is the sin that resides in our hearts. That is why the message of the gospel is that God transforms our lives by transforming our hearts.” And again, “Even though life is hard, it is not the hardships that cause us to respond as we do. Our responses are shaped by the thoughts and motives of our hearts (Heb. 4:12).” The authors also remind us that in Christ and His gospel, we have a new record, a new power and are set free from the enslaving power of sin. They write, “With the new record and new power we have received, we are set free from the enslaving power of sin and the condemnation of the law. We are dead to the world and we have power over the Evil One who tempts us with worldly Christ-replacements. We don’t have to be controlled by them anymore! We are now free to live, act, think, and believe in new and surprising ways.” The authors also remind us of our marriage or union to Christ, and how God designed heart change to take place in the context of community (the church) – something we conveniently ignore in our mostly individualistic culture! The latter part of the book focuses on four themes: Heat, Thorns, Cross and Fruit. “Heat” is the pressures of everyday life and includes blessings, temptations and difficulties. “Thorns” is the ungodly response to the daily situations of life. “Cross” is the resources available in Christ to every believer. “Fruit” is the Godly response to the situation resulting from God’s work in the heart. With sharp Biblical insight and relevant everyday stories, the authors present a mirror in front of our souls, so that we see the hope & power of Christ and His gospel for everyday living. Heart change is possible! Read this book and apply its truths! I hope to do the same.
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Reviewed in the United States on September 2, 2016
★★★★★ 5
Lasting change
Format: Paperback
What does it take for lasting change to take root in your life? This is the question Timothy Lane and Paul Tripp tackle in the book. Both are biblical counselors and faculty at CCEF (Christian Counseling and Education Foundation - maybe the best biblical counseling group out there). Paul Tripp is also a pastor at Tenth Pres in Philadelphia.
The authors believe that in the lives of many believers (and a whole lot of churches) there is a "gospel gap." Many Christians are ineffective and unproductive because they see the good news of the gospel as the means of cleansing their past sins, and the way to be assured of a future hope. But they forget the power of the gospel to change their life in the hear and now.
Lane and Tripp use Jeremiah 17:5-10 as the source for their dominant metaphor for life change. This passage gives us a view of life that involves four elements.
1. Heat - This is the person's situation in daily life, with difficulties, blessings, and temptations.
2. Thorns - This is the person's ungodly response to the situation. It includes behavior, the heart driving the behavior, and the consequences that result.
3. Cross - This focuses on the presence of God in his redemptive glory and love. Through Christ, he brings comfort, cleansing, and the power to change.
4. Fruit - This is the person's new godly response to the situation resulting from God's power at work in the heart. It includes behavior, the heart renewed by grace, and the harvest of consequences that follow.
Each chapter has large sections of careful explanation of biblical passages. Lane and Tripp also use real life examples from their counseling and church ministry to illustrate the point. It's both theological and practical. It's a great book for those who do counseling, or even just general discipleship. It's a good book for those wanting to better understand the process of sanctification, and how they themselves can grow. I'm looking forward to looking through the curriculum based on the book, and see if we can find a venue to use it in our church.
Highly recommended!
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Reviewed in the United States on June 19, 2010