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Description
Human TLR2 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: Saliva 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).
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: Saliva 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: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 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 uses 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 Toll-Like Receptor 2 (TLR2) 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 Toll-Like Receptor 2 (TLR2) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Toll Like Receptor 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Toll-like receptor 2, also known as TLR2, is a protein encoded by the TLR2 gene. TLR2, also known as CD282 (cluster of differentiation 282), plays a role in the immune system. It is a membrane protein and a receptor expressed on the surface of certain cells that recognizes foreign substances and transmits appropriate signals to cells of the immune system. The protein encoded by this gene is a member of the Toll-like receptor (TLR) family and plays a fundamental role in pathogen recognition and activation of innate immunity. TLRs are highly conserved from fruit flies to humans, sharing structural and functional similarities. They recognize pathogen-associated molecular patterns (PAMPs) expressed on infectious pathogens and mediate the production of cytokines required for effective immune development. Different TLRs exhibit distinct expression patterns. This gene is most abundantly expressed in peripheral blood leukocytes, mediating host responses to Gram-positive bacteria and yeast by stimulating NF-κB. | |||||||||||||||||||||||||||||||||
| 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 | Saliva, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.1 ★★★★★
Based on 25 reviews
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Product Reviews
★★★★★ 4
Good but not as good
Format: Kindle
Freedom From Fear #1 was one of the most interesting and informational books I've ever read. Part Two was interesting but I did not learn much more about the war than I had known before other than the fact that, while Japan lost the war, Anglo-Saxons are no longer in control in the Far East. I somewhat disagree with his ending ideas about the "good" war, that it wasn't so "good" after all in it's outcome. However, those who fought in it and their families would probably disagree.
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Reviewed in the United States on May 22, 2014
★★★★★ 5
The Innovator's DNA - Disruptive Research - Disruptive Writing
A Politically Correct Status Quo
It is politically correct in management circles to say that you are "results oriented" or that you "drive for results" in your organization.
The status quo in business schools is to indoctrinate students in the delivery skills of analyzing, planning, detail-oriented implementing, and disciplined executing.
This book and the research upon which it is based disrupts that politically correct status quo.
Clayton Christensen has spent close to two decades creating the research, conceptual, and application foundation of the disruptive innovation body of knowledge. He has been working for more than 8 years with Jeff Dyer and Hal Gregersen, both gifted researchers, teachers, and consultants in their own right, on this project. These guys are a disruptive "dream team" of contributors.
This book articulates an extension of the disruptive innovation body of knowledge that clearly describes an individual profile of the disruptive innovator and an organizational profile of an organization that makes disruptive innovation happen.
So what makes this book disruptive?
The first thing is timing. It arrives on the scene at a time when innovation is one of the most critical components of a solution to our global financial and organizational mess. If we are to get out of our morass of debt and sluggish growth and respond to the continually emerging challenges of a burgeoning global society it will ride on the backs and wings of innovation. The status quo must be disrupted for us to survive and thrive!
Second is the audacity of the core models. The authors claim that innovation can be learned at both the individual and organizational level. Individuals can increase their ability to discover (Discovery Quotient - DQ) and learn to be more innovative. They cite the four specific behavioral skills of asking questions, engaging in observations, networking with people who have a different point of view, and experimenting to figure out what can work as the common elements of what innovators do. They also identify the cognitive skill of associational thinking, the ability to find connections between ideas that do not seem to be related to each other, as the connection between the behavioral skills and the generation of ideas. They extend their claim that the innovation competency can be learned to the organizational domain by saying that organizations can become more innovative through developing and leading people, designing and implementing processes, and advocating and living by philosophies that support innovation. These two arguments stand in stark contrast to the beliefs and practices of a vast majority of leaders and institutions.
(For a diagram of the Model see [...])
'And all of this is built upon the third source of disruption: research. Their work is based on well-founded research into the "DNA" of the world's leading innovators and the world's most innovative organizations. The authors conducted nearly 100 interviews of world class innovators and their colleagues to get at the heart of what innovators do. They also interviewed and surveyed executives who are not innovators. (Their survey data base has over 5000 respondents in it.) So they have been able to compare and contrast the two populations to more clearly see what it takes to effectively innovate.
They have also done research on business results attributable to innovation. Collaborating with HOLT (a division of Credit Suisse) they were able to craft a measurement called the "innovation premium." This measure identifies if an organization's market capitalization can be accounted for by existing cash flows or if there is an innovation influence on the stock price. By using this measure, they have been able to clearly and objectively identify which organizations are benefiting from innovation.
Yet to Explore
The tension in the balance of influence and power between the leaders with predominantly "Discovery" or "Delivery" mindsets is an area that has yet to be explored. If the premises of this book are sound, and I believe they are, we need to figure out how to manage that tension and balance in order to generate, incubate, and strengthen innovative ideas as we bring them to full fruition in the marketplace. Great ideas that are not delivered upon are simply recreational pursuits that do not build great people, great institutions, and great societies. So there is work yet to do.
Invest Your Time and Effort
This book makes a significant contribution to both the disruptive innovation body of knowledge and the evolving body of practice on innovating disruptively. It is well worth reading, pondering, and acting upon.
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Reviewed in the United States on August 18, 2011
★★★★★ 5
Inspiring and well-written
This is a very interesting book written by some Harvard profs. They did a large national survey of innovative businesses and their leaders. The book posits that innovative people follow five skills: associating, questioning, observing, networking, and experimenting. These skills can be found at the individual or organizational level. The idea is that most people have these skills in their DNA and can bring them out with some practice. There are a lot of interesting and inspiring examples like Steve Jobs and Jeff Bezos. Although this book seems like a self-help type book with a lot of hype, it has an academic underpinning. Any organization that is interested in promoting innovation could benefit from encouraging these 5 skills. If you are interested in innovation or creativity in business or any organization that produces something, you will like this book. The books is a little distracting to read because it has sidebars all through it giving interesting examples that break up reading concentration. Aside from that, it is a well-written book that is easy and enjoyable to read. I enjoyed the book greatly and found it to be inspiring.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 5, 2015
★★★★★ 4
Great read and research. But what about daily application for regular people?
The research piece behind this book might be the next thing I read, as I'm intrigued by the academic rigor applied.
The reveal and living examples of the five skills - questioning, networking, experimenting, observing and associating - are tangible and approachable given their articulation through well-known and highly visible entrepreneurs running innovative companies. There's much to be gleaned by looking at the way these people behave and, even through simple emulation, enhancing one's own skills.
My only real disappointment with the book is its limited approach to practical, daily application for those not yet at the top of the tree. It's rather a different kettle of fish for the innovation-minded, but stuck in bureaucracy, worker who wants to make things better, is still motivated, and hasn't been crushed by the machine. How does that person actively innovate? And, in some cases, get away with it? This book (or an accompanying volume) focussing on daily, in-work, innovation would be useful.
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Reviewed in the United States on May 23, 2013
★★★★★ 5
A Favorite Book on Innovation
Format: Hardcover
Very well written and enough stories to help the true content stick. This is a favorite book of mine and has lead to interesting conversations to boot.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 29, 2025
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