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Description
Human HDL2 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: 1. 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: 1. 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. 2. 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. 3. 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. 4. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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 high-density lipoprotein 2 (HDL2) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of high-density lipoprotein 2 (HDL2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human high-density lipoprotein 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | High-density lipoprotein (HDL) is primarily synthesized in the liver and small intestine. Nascent HDL synthesized in the liver is primarily composed of phospholipids and ApoAⅠ. Under the action of LCAT, free cholesterol is converted into cholesterol esters, and the lipoprotein is transformed into mature globular HDL3. This is then converted into HDL2 by LPL. High-density lipoprotein 2 (HDL2) is an autosomal dominant inherited disorder that manifests as various movement disorders, including chorea and dystonia. | |||||||||||||||||||||||||||||||||
| 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 homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.3 ★★★★★
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Product Reviews
★★★★★ 5
Great value, cheaper than local.
Style: Full Synthetic High Mileage, Size: 5 qt (Pack of 1), Configuration: 0W-20
Good value, fast shipping, Valvoline quality.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 16, 2026
★★★★★ 4
Good buy
Style: Full Synthetic High Mileage, Size: 5 qt (Pack of 1), Configuration: 5W-20
Run of the mill oil but for a great price
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Reviewed in the United States on March 27, 2026
★★★★★ 5
The Excellence of Motion Preserved
Style: Full Synthetic High Mileage, Size: 1 qt (Pack of 1), Configuration: 5W-30
In the pursuit of the ideal, where reason governs and the forms of all things aspire to perfection, the Valvoline Full Synthetic High Mileage with MaxLife Technology 5W-30 Motor Oil presents itself as a manifestation of virtue within the mechanical realm. It is not merely oil, but a substance designed with foresight, sustaining the engine as the soul sustains the body. The viscosity is measured, neither excessive nor deficient, allowing parts to move in harmonious accord, reducing friction and preserving integrity.
One observes that engines treated with this oil respond with steadiness and endurance, as if guided by a rational principle, minimizing wear and extending life in a manner that reflects the pursuit of the Good. I grant it five stars, for it exemplifies a balance between strength and refinement, a practical embodiment of foresight, wisdom, and care—ensuring that motion, that vital energy, continues undisturbed, much as a well-ordered soul achieves its fullest expression through the contemplation of virtue.
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Reviewed in the United States on December 13, 2025
★★★★★ 5
Extraordinary resource
Format: Paperback, Format: Paperback
I am a Cultural History Interpreter in SC. Working at a plantation historic site to bring suppressed history to light is challenging. Prof Sinha's book gives us easily accessible documentation to counter the "Lost Cause" devotees who appear on the site almost daily.
Her writing style is clear and lucid, a trait for which I am extremely grateful. The site is including this volume in our staff library. For those just entering the field of Public History, it is indispensable. For the rest of it is a very valuable resource. Highly recommended!
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Reviewed in the United States on May 8, 2019
★★★★★ 4
An important contribution
The historiography of secession is a complex one. For much of the last century there had been a tendency for historians to underplay the importance of slavery as a cause of the American civil war. Certaintly neo-Confederate apologists have sought to euphemize the cause of the conflict to an issue over tariffs, to matters of states rights, or to the "extremism" of the abolitionists. It is quite clear that these excuses will not survive a reading of this book. Sinha clearly shows, in her examination of South Carolina secessionism from nullifaction to fort Sumter, that slavery was the essence of its concerns. To show this she looks at the nullification crisis, the Mexican war, the Compromise of 1850, the South Carolinian movement to reopen the slave trade, and the secession crisis, based on exhaustive research of no less than 137 sets of private papers and diaries.
But Sinha wishes not simply to refute the academically unimportant group of neo-Calhounites. She wishes to argue something broader. The South Carolinian defense of slavery was not, as many serious historians suggest today, simply the working out of the Southern American view of liberty. Increasingly, Sinha argues, South Carolina pro-slavery thought was not the expression of Southern Republicanism, but increasingly its very negation. It was not a coincidence that secessionism was strongest in South Carolina, the only state by 1832 where presidential electors and the governor were not popularly elected, where the legislature was crudely malapportioned, and where local offices were limited by the state government. It was also not a coincidence that slaves were a majority of South Carolinians, and slaveholders nearly a majority of South Carolinian whites. And it certainly was not a coincidence that non-slaveholders were noticeably less enthusiastic for nullification, secession in 1851 and secession in 1861.
But although Southern nationalist discourse was clearly elitist and pro-slavery, does Sinha show that it was counter-revolutionary? A certain opposition to democracy was evident after all in the many, perhaps most, of the founding fathers. But as Sinha points out leading Carolinians like Calhoun, Senator James Chesnut and the creepy, incestuous James Hammond all sneered at the Declaration of Independence. She quotes one bravado warping PatricK Henry to declare "Give me Slavery or give me death." Notwithstanding the views of some historians to the contrary the South Carolinians criticized the North less for its oppression of wage laborers than the possiblity that those laborers could vote themselves into power. They did not condemn Lincoln as an intolerant Protestant but as a dangerous socialist and feminist. Moreover, they were not slow to raise the Nativist card against the immigrants who were bolstering the North's population. Calhoun's idea of a concurrent majority was not a thoughtful protection of minority rights, but a way to prevent one minority, his own, from ever being outvoted. Once the Confederacy was set up the elite dispensed with political parties. Looking at South Carolina they also began to dispense with competitive elections, while its ruthless elite certainly did not act sentimentally (or even decently) towards opinions on slavery.
In conclusion there have been many frauds and bullies in American political life: the Nixons, the Hoovers, the McCarthys, the Tillmans and the Bilbos. But much of their malignancy was purely personal and they never threatened the core ideals of the republic. Calhoun was different, very different. Extremely intelligent, he was also utterly principled, and absolutely ruthless in carrying out that one principle. The problem was that the principle, despite all the complications of honor and paternalism, was slavery. More so than anyone else, Calhoun was the greatest enemy of liberty and freedom the United States ever had. Sinha's book is an important contribution to understanding that.
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Reviewed in the United States on December 29, 2000
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