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Description
Human EPB42 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 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 an Erythrocyte Membrane Protein Band 4.2 (EPB42) 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 Erythrocyte Membrane Protein Band 4.2 (EPB42) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Erythrocyte Membrane Protein Band 4.2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Erythrocyte membrane protein band 4.2, also known as EPB42, is a protein encoded by the EPB42 gene. It is part of the erythrocyte cytoskeleton. It is an ATP-binding protein that regulates the binding of band 3 to acetylcholine. It may play a role in regulating erythrocyte shape and mechanical properties. Mutations in this gene are associated with recessive spherocytosis and recessive hereditary hemolytic anemia. | |||||||||||||||||||||||||||||||||
| 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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4.7 ★★★★★
Based on 13 reviews
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Product Reviews
★★★★★ 5
Definitely buying again.
Style: High Mileage 150K, Size: 5 qt (Pack of 1), Configuration: 5W-20
In MY experience, this has been fantastic, spouses car is a 200k+ mileage Kia shortage, which I mean, it’s always burned oil, even the manual said normal operation can burn 1qt per 1,000 miles. Either way, i was adding oil a lot. At least 2 quarts a month or more. After running this oil, I have only added 1.5 quarts in over 2 months. Results obviously not typical, but in my experience, this stuff works. I also use the restore and protect Valvoline on my car, with great results. Scoped it after a few full oil changes and engine looks clean. Great value for what it potentially will do.
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Reviewed in the United States on March 2, 2026
★★★★★ 4
Kitties love it:
Style: High Mileage 150K, Size: 5 qt (Pack of 1), Configuration: 5W-30
Great product:
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Reviewed in the United States on April 23, 2026
★★★★★ 5
Engaging, though-provoking sweep that will provide you with regarding this time period
Format: Hardcover
Alan Taylor is a writer who excels at contextualizing the complexity of history by creating a sort of ancestral snapshot of each person and event and placing them on a family tree, showing both their relationships to one another and to their time. This approach increases readers’ abilities to build those understandings on their own in other readings, about other times. That’s cool. In this book, he upends a more static understanding of North and South and provides a kaleidoscope of complexity with regards to individuals and social groups from regions both within and outside of our borders.
In this book, Alan Taylor displays his unique brilliance at making legible the complex interplay of extremely diverse international, national, and factional agendas, political aspirations, people’s attachment to their political and social worldviews, economic aspirations, their bluster, their denial, and their honest – if not always successful – efforts. Quoting from a mind-bogglingly large reading list of academic sources, newspapers, diaries, and other historical documents, he brings people back to life in such a way that you could mentally animate what role these historical figures would play today on the world stage or even in a more intimate setting of your own office politics. He makes the complexity and uncertainty decipherable so that we can think about it, argue about it, and explore it just as we would events with which we are familiar today.
A true love of history and our understanding of humanity at present are not served by infatuation with imagined, polished heroes but by complex accounts and considerations of character, influences, dreams, successes, and failures that reveal how these elements are the common denominators in all lives and across all times. Taylor does this superbly for figures North, South, enslaved, free, freed Blacks, embittered whites, Mexican, Spanish, Canadian, British, French, and Indigenous. He juxtaposes Maximilian’s wife, Carlota, sister of Leopold II, who placed faith in herself and in her husband to transform Mexico through better monarchy, with the far more egalitarian Benito Juárez, who ultimately subordinated the lives of the indigenous people in capitulating to a rising oligarchy of American investors who could rebuild Mexico. Both Carlota and Juarez are driven to varying degrees of madness by the results of their efforts.
We see members of the former Confederacy who rue their violent support for the perverse and cruel institution of slavery once the war is over, alongside others who will stop at nothing to bring back the old order. And we see Northerners, who in wartime decried slavery with a furious ardor, eventually languishing in their duty to their fellows after the war was over. There are warriors for justice, warriors for oppression, realists, capitulators, power brokers, and pawns. Even the best, who are not depleted of passionate intensity for doing right, must contend with an ecosystem of others’ dreams and aspirations, which all too often run afoul of the righteous. In the end, we may be judged by others and by ourselves for what we’ve wished for: either peace and fairness or war and acquisition at any price.
The book serves as a reminder to plant the right seeds and dream the right dreams…for everybody’s children. Because when the harshest frost melts away, something new will grow.
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Reviewed in the United States on July 3, 2024
★★★★★ 5
Carefully Researched Gives Insight To 19Th Century Occurances of: American, Canada, and Mexico!
Format: Hardcover
This book is a treasure as it covers not only the American Civil War but what intricate details are behind it and more, in addition covers the same eras for the Dominion of Canada, and French take over of Mexico along with the factors leading to "Cinco De Mayo," and more. As an avid reader of American History also as a amature historian this book is carefully detailed and gives insight to the racial and political beliefs at the life and times of the 19Th. Century. It deserves a place on your bookshelf and/or library. In these contemporary times, I am still more than pleased the the border frontiers between the Republic of Canada and United States of America remain the: "Longest Undefended Borders" in the entire globe.
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Reviewed in the United States on May 5, 2025
★★★★★ 4
Another Thought Provoking Book
Format: Kindle
Having read Professor Taylor's American Republics I greatly anticipated this volume in his series. The examination of both the Canadian and Mexican stories in this book along with the American Civil War helps provide context to the traditional narrative. I find his approach useful as it shows how the interactions between the US and its neighboring nations evolved. I'm hoping he continues the series
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Reviewed in the United States on September 1, 2024
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