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Description
Mouse TNC 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: 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: 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. 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. 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. 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 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 tenascin C (TNC) 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 tenascin C (TNC) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Tenascin C ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Tenascin C (TNC) is an oligoglycoprotein composed of a single polypeptide with a molecular weight ranging from 180 to approximately 300 kDa, encoded by the TNC gene. Proteins in the tenascin family share a similar structural pattern. These similar modules include heptagonal repeats, EGF-like repeats, a fibronectin type III domain, and a C-terminal globular domain shared with fibrinogen. These protein modules are arranged in a row like beads, resulting in long, extended molecules. At the N-terminus, each tenascin has an oligomerization domain, which, in the case of TN-C, forms a hexamer. In addition to eight constant repeats, nine additional repeats exist that are subject to alternative splicing. During development, disease, or injury, it is expressed in the extracellular matrix of various tissues and in limited neurogenic regions of the central nervous system. In the embryo, it is produced by migrating cells such as the neural crest. It is also abundant in developing tendon, bone, and cartilage. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenates and other biological fluids |
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4.5 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 4
Foes at the Top Table
Format: Kindle
Those of us who studied economics in the 60s grew up on Keynes. This book provides a fascinating picture of the great man in action. And an equally fascinating picture of the Lend Lease negotiations and then the US hard line at Bretton Woods. Behind this hard line was Harry Woods, of Lithuanian emigre stock, who clawed his way by hard work and intelligence to negotiating prominence in the US Treasury. And who was a Soviet agent of influence. Well written, lucid, and remarkably interesting.
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Reviewed in the United States on May 19, 2013
★★★★★ 5
A distant mirror of our current problems
Format: Hardcover
The title of this excellent book accurately describes its contents:it is about a battle fought to define a new world order, that which was emerging from the ashes of World War II. The book also conveys the messy complexity of such a historical process--how individual characters interpreted the events around them, realized that they were giving shape to a radically new future, tried to take advantage of them to advance their own personal and national interests, and succeeded in accordance with their intelligence, the cunning of their argumentation, and, above all, the shifts in the real power that supported them. Masterly, Benn Steil makes the reader feel how Keynes and White gradually reached an unspoken and unrecognized agreement regarding the shape that the new world would have, and then fought to gain advantage in that new world--Keynes trying to keep the British Empire paramount in the world order, now based not on the Royal Navy but on Britain's alliance with the United States, the emerging superpower, and White asserting the unimpeded power of the United States. Focusing on one crucial aspect of the new order, money, Steil is able to reenact the human drama of the transfer of world power from Britain to the United States in all orders of life. It is an excellent history book.
The book, however, goes beyond history as the narration and understanding of past events. When reading it, there is an eerie feeling that you are reading about current events. The process that led to Bretton Woods started thirty years before, with World War I and the end of the classical gold standard. When the war ended, a new monetary system was created, which was called the gold exchange standard. It resembled but emasculated the power of the old gold standard to keep monetary order in the world at large. This new system gave central banks the power to create money independently of the international consequences of doing it. With time, central banks abused this power, created a boom in the 1920s and then a depression in the 1930s. Bretton Woods was convened to reintroduce order in the monetary world. Like the gold standard of old, the new system created there was tied to gold in an effort to ensure stability. Yet, it also allowed central banks freedom to create money under certain circumstances. As it happened in the 1920s and 1930s, central banks abused their power, blew up the international system (in this case the Bretton Woods system) and then led the world into a series of booms and busts that has not ended as yet. A new monetary order will be needed to avoid worldwide inflation and protracted recessions. To understand the issues that will be crucial to give shape to this new monetary order it will be necessary to revisit the making of Bretton Woods in detail. There is no better way to understand these issues that Ben Steil's The Battle of Bretton Woods. Thus, in addition to being an excellent history book, it is also an excellent book about current events.
Full disclosure: I wrote a previous book with Benn Steil: Money, Markets and Sovereignty (Yale University Press, 2009).
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Reviewed in the United States on May 28, 2013
★★★★★ 5
The Battle of Bretton Woods
Format: Hardcover
From the growing reliance upon international finance and the devastating repercussions of two World Wars, Steil weaves together an important narrative that tells the story of America's rise to the world stage as a major power. Britain's reign of dominance comes to an abrupt end under the weight of the Second World War and the dependence of their territories. Reliance on foreign aid and mounting debt put Britain in a precarious situation for which the United States capitalized on to secure its place as the dominant world power. Through the ideas and experiences of two brilliant economists, Harry White and John Keynes, were guided through the creation and implementation of an economic solution to remedy, and further amalgamate the global financial framework. At Bretton Woods, White and Keynes promote slightly different plans that form the International Monetary Fund and World Bank; organizations designed to monitor, stabilize, and assist international finance. To Britain, and much of the world's chagrin, the organizations are formed in a manner that benefited the United States post-World War Two position as a creditor nation. Dollar dominance in the newly designed financial markets promotes short-term growth for the United States. However, financial mismanagement and over-extension soon lead the U.S. down a path of monetary hardship that ultimately results in our current situation as debtor nations (much like Britain was when the story began). The author ends the narrative by chronicling the effects of Bretton Woods on the United States, Britain, and international finance from the mid-twentieth century to the present. This book tells the remarkable story of America's rise to power through a financial lens. Steil is a wonderful writer who describes complex ideas of monetary policy, international economics, and currency manipulation in such a manner that is easy to understand and leaves the reader wanting more.
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Reviewed in the United States on August 16, 2013
★★★★★ 5
A concise, readable history of European developments prior to W.W. i.
Format: Paperback
This is part of a 4 volume series on (primarily) European history covering the development of the French and Industrial revolution(s) with particular emphasis on the Belle Epoque that marked the end of nearly 100 years of peace among the European Powers. It covers a variety of topics ranging from the emergence of the working classes; the role of the middle class; industrial capitalism; nationalism; the sciences and the arts; and, even a chapter on the "New Woman". Quite encompassing in its treatment and its analysis of the a period that serves as a background to the twentieth century.
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Reviewed in the United States on August 3, 2018
★★★★★ 5
Un comentario
Format: Paperback
El mundo burgués generó desarrollos muy importantes para los habitantes de Europa Occidental particularmente; incrementó de manera importante los niveles de vida de una porción significativa de la sociedad, introdujo innovaciones claras en las ciencias, en lo educativo, en el comercio global y en el intercambio entre diferentes culturas. Mejoró la productividad, las comunicaciones, la alimentación......Pero también incorporó consigo tendencias negativas y difíciles que cambiaron para siempre la interrelación entre pares al crearse una diferencia significativa de clases, la alta, burguesa y la trabajadora y proletaria, continuó la discriminación sobre la campesina, estableció el concepto forzado de nacionalismo, transformó a los países mas desarrollados en imperios con ansias de crecimiento y de dominio de los menos preparados, se evidenció la explotación del tercer mundo, se inició el armamentismo y la instigación a la guerra que culminó con la desastrosa confrontación de 1914. El mundo, a partir de ese momento no volverá a ser igual.
Todos estos conceptos, esta historia, están amenamente descritos por el autor, con un gran conocimiento de la época y con una inteligente asociación de los acontecimientos para hacer entender al lector sus grandes transformaciones e implicaciones.
Como los otros tres libros de la serie, lo recomiendo ampliamente. Permite entender todo la evolución de nuestra civilización en los dos últimos siglos así como la influencia generada por los grandes imperios de la época y sus pensadores
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Reviewed in the United States on November 28, 2011