Pay in installments of $46.03 with
,
and
Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Aug 18 - Aug 23
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
Description
Rat 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 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. |
|||||||||||||||||||||||||||||||||
| 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 peroxidase (HRP) catalysis 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 | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat High-density lipoprotein 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
|
|||||||||||||||||||||||||||||||||
| 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 ApoAⅠ. 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 by 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. |
|||||||||||||||||||||||||||||||||
| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
Shipping Notes
- Free Standard Shipping on $100+ Orders to the USA.
- Except Preorder products are shipped in 48 hours.
- Delivery to the USA:
- Standard Shipping : 3-10 business days
- If time is of the essence, please consider selecting expedited delivery for faster service.
Exchange/Return Notes
- We offer a 30-day return/exchange service after receiving.
- Final sale items are not eligible for returns or exchanges.
- To process your return/exchange, please contact us at [email protected]
- Please click here for more details>>> Return & Exchange Policy
4.6 ★★★★★
Based on 23 reviews
Sort
Product Reviews
★★★★★ 5
"Durable Fun: Pweituoet Heavy Duty Squeaky Dog Balls for Aggressive Chewers"
Size: 4.5"
Product Review:
The Pweituoet 4.5” Heavy Duty Squeaky Dog Balls have been a hit with my dog, proving to be both durable and entertaining.
Durability:
These dog balls are impressively tough. My dog is an aggressive chewer, and most toys don’t last long, but these balls have held up exceptionally well. The heavy-duty material and spiked design are perfect for dogs who love to chew, providing both durability and safety.
Entertainment:
The built-in squeaker adds an extra layer of fun. It captures my dog’s attention immediately, and the squeak keeps him engaged for longer play sessions. The spikes also make the ball easy for him to grip and carry around.
Size:
The 4.5” size is ideal for medium to large dogs. It’s large enough to avoid being a choking hazard but still easy for my dog to pick up and play with. The size and weight are just right for games of fetch, too.
Value:
Given the durability and how much my dog enjoys these balls, they offer great value for the price. They’ve lasted much longer than other toys we’ve tried, making them a worthwhile investment for any dog owner with a tough chewer.
Overall Verdict:
The Pweituoet Heavy Duty Squeaky Dog Balls are an excellent choice for medium to large dogs, especially those who are aggressive chewers. They’re durable, fun, and well-sized, making them a great addition to your dog’s toy collection. Highly recommended!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 10, 2024
★★★★★ 5
Actually stands up to use
Size: 4.5", Size: 4.5"
I have a pitbull who is a toy destroyer. All those toys, even the ones that say tough enough for destroyers, never hold up. THESE BALLS ARE AWESOME! I bought the 2 pack of the 4.5 inch balls and they lasted a year. I literally am writing this review after ordering a new set. These are well worth the money and Ghost (my dog) loves them. Hes a 103lbs pitbull and his life mission is cuddles and destroying his toys... and these lasting a year of everyday use is friggin awesome. Both toys failed in the same way eventually. They gave out in a circle around the "squeker".
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 14, 2024
★★★★★ 3
Neutral
Size: 4.5"
She chewed it up in less than 10 minutes. This is not for heavy chewers but dogs that like to actually play with their toys it would be great!!! I gave it a 3 because it worked for my other dog who likes to play🤘
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 8, 2026
★★★★★ 5
Our German Sheppard loves these
Size: Medium
These cost a bit more than tennis balls, but they are so much nicer and longer lasting. For starters, they stay cleaner than tennis balls because they’re smooth rubber. Dirt won’t build up on them and if anything does stick, like grass or soil, it falls off once the dog slobber dries. They’re also thick, so they don’t fall apart or blow out like a normal tennis ball does in our dog’s jaws after 30 seconds. Our GS chomps on these like crazy and the only damage they’ve suffered is a crack that developed from the edge of the hole, but the crack is growing very slowly and none of these balls have totally failed yet. The balls do whistle when thrown ant high speed and that may help a dog track and locate it, but I’m not sure. Our neighbors hear the whistling too so it’s far from silent. Lastly the orange ball is easy to locate out in our yard, but the dark blue practically disappears.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 6, 2025
★★★★★ 5
Great for smaller dogs
Size: Small
These two balls are perfect for the smaller mouthed dog that loves to play fetch.
These balls are not only super durable (lots of teeth biting), but float in the baby pool we use for our miniature dachshunds.
The value here is much better than you’d find anywhere else. The noise, if bitten hard enough, was “low” at best. Easy to spot/find if overthrown. Will definitely buy again once these are in bad repair; so far, so good-love these for my fur babies!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 8, 2024
recommand products
FPF Intank EFI Fuel Pump w/ Tank Seal For Polaris Ranger 700 / XP / Crew 2013-2018, Replaces 3203641
79.99
Archie Super Digest Pack
128.00
FPF T35 Intank Fuel Pump Honda NCH50 / 2AC 2013-2018
79.99
Cusco Street Zero-A 08-12 Subaru GRB/GVB STi 40-Way Adj Front/Rear Pillow Upper Mounts Coilovers - 692 61N CP
945.00
FPF T35 Intank Fuel Pump With Regulator And Strainer For Kawasaki Vulcan 900 2008-2020, Replaces 49040-0022
89.99