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Description
Mouse HDL3 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 3 (HDL3) 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 high-density lipoprotein 3 (HDL3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse high-density lipoprotein 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | High-density lipoprotein (HDL), a serum protein, is a complex lipoprotein composed of lipids, proteins, and the regulatory factors they carry. It is also known as a1 lipoprotein. HDL is primarily synthesized in the liver and small intestine. Nascent HDL synthesized in the liver is primarily composed of phospholipids and ApoA I. Under the action of LCAT, free cholesterol is converted into cholesterol esters, and the lipoprotein becomes mature globular HDL3, which is then converted into HDL2 through the action of LPL. LCAT completes the conversion of nascent discoidal HDL into HDL3 and HDL2 through transesterification, reducing the concentration of free cholesterol in plasma HDL, establishing a concentration gradient for cholesterol to flow from cell membranes to plasma lipoproteins, and reducing tissue cholesterol deposition. | |||||||||||||||||||||||||||||||||
| 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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4.0 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 5
Metamucil 4-in-1 Psyllium Husk GLP-1 Friendly Fiber Supplement, Sugar-Free, 180 teaspoons, Orange
Size: 2.3 Pound (Pack of 1), Style: Sugar Free 180 count
I started taking Metamucil after my doctor recommended boosting my daily fiber intake, and I genuinely wish I had started sooner. This sugar-free orange flavor has become a non-negotiable part of my morning routine.
First, the taste — for a sugar-free supplement, it's surprisingly pleasant. The orange flavor is light and natural, not artificial or overpowering. Mixed into a glass of cold water, it goes down easily without that chalky, gritty aftertaste I dreaded from other fiber products I've tried in the past.
The 4-in-1 benefits are the real selling point for me. I've noticed a genuine difference in my digestive regularity, I feel fuller between meals (which has helped me make better snack choices), and my energy levels feel more stable throughout the day. As someone on a GLP-1 medication, I was thrilled to find a fiber supplement specifically formulated to complement that — it fills a real gap in the market.
The 180-teaspoon count is outstanding value. This container lasts a long time, which means fewer reorders and a lower cost per serving than most comparable products. The scoop is easy to use and the powder dissolves well with a quick stir — no clumping.
My digestion has genuinely improved since making this a daily habit. Less bloating, more consistency, and I just feel better overall. It's one of those quiet little wellness wins that adds up over time.
If you're on the fence, just try it. Simple, effective, great value — this is now a permanent fixture in my pantry. Cannot recommend it highly enough.
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Reviewed in the United States on May 25, 2026
★★★★★ 5
Does its job very well
Size: 2.3 Pound (Pack of 1), Style: Sugar Free 180 count
Been using Metamucil for years. Because it is a quality product that dissolves well in water, tastes fine, and a very easy way to stay regular. Considering what it does for my system, it is a cost efficient way to get more fiber in my diet with no side effects.
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Reviewed in the United States on April 14, 2026
★★★★★ 5
Excellent product!
Size: 2.3 Pound (Pack of 1), Style: Sugar Free 180 count
I held on to this product for months before trying it. Wow! Not sure what took me so long, but I am so impressed. First of all, it is easy to drink - just do it quickly. The taste is surprisingly refreshing.
Even more important, it works. If you have IBS or other digestive issues, you may be pleasantly surprised at how it helps these issues. I just regret I didn’t start using it years ago. I take at least two drinks a day, sometimes only one.
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Reviewed in the United States on May 27, 2026
★★★★★ 4
Works well and easy to add to a daily routine
Size: 2.3 Pound (Pack of 1), Style: Sugar Free 180 count
This Metamucil works well overall. It mixes pretty easily, the orange flavor is decent, and it’s been a simple way to add more fiber into my daily routine.
I like that it’s sugar-free and uses psyllium husk, so it feels like a practical option for digestive health without adding extra sugar. The texture is what you’d expect from a fiber powder, so you do need to drink it fairly quickly after mixing.
Overall, it does what I bought it for. Taking off one star only because it’s still a fiber drink, so the taste and texture may not be for everyone, but it’s a solid product.
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Reviewed in the United States on May 28, 2026
★★★★★ 5
This really works and tastes great
Size: 2.3 Pound (Pack of 1), Style: Sugar Free 180 count
Top notch quality
The only brand I buy !!!!!!
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Reviewed in the United States on May 4, 2026