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Description
Human H+/peptide transporter 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 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. |
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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 capture antibody against Solute carrier family 15 (H+/peptide transporter), member 2 (HPEPT2). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Solute carrier family 15 (H+/peptide transporter), member 2 (HPEPT2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Solute carrier family 15 , member 2 (HPEPT2) ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Hydrogen ion/peptide transporter 2 (HPEPT2), also known as solute carrier family 15 member 2, is an enzyme encoded by the SLC15A2 gene. It is a proton-coupled amino acid transporter that transports oligopeptides of 2 to 4 amino acids, with a preference for dipeptides. It transports the dipeptide-like aminopeptidase inhibitor bestatin, the aminocephalosporin antibiotic cefadroxil, and carnosine. It participates in innate immunity by facilitating the detection of microbial pathogens by NOD-like receptors. It may function by mediating the transport of bacterial peptidoglycans across the plasma membrane, catalyzing the transport of certain bacterial peptidoglycans, such as muramyl dipeptide (MDP), a NOD2 ligand. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.2 ★★★★★
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Product Reviews
★★★★★ 5
Versatile water bottle_ great ice cold retention
Color: Strawberry Fields, Size: 24 Ounces, Color: Strawberry Fields, Size: 24 Ounces
There are a lot of stainless steel bottles out there but the quality of Owala stands out by far. Starting with the both drink options of sip or chug, mega cute color combinations, ergonomic grip, cold retention is great and different size to fit everyone of your family's thirst. Easy to carry it with its handle. It's almost a month that I've been using it and no spills on my backpack. Defo buying more on the future, just to change the color with the Seasons. Rubber Boot is not included but make sure to add it to your cart...is soooo worth it!
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Reviewed in the United States on May 12, 2026
★★★★★ 5
Water bottle
Color: Shy Marshmallow, Size: 24 Ounces
My son had to have this water bottle and at first I was like what's all the hype about but I know now bc it literally the best water bottle, it super easy to clean, its versatile, it keeps your drink cold all day, the lid is amazing bc there's no leakage at all and you can use a straw or the drinking part for your mouth, great value for the money
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Reviewed in the United States on May 24, 2026
★★★★★ 5
great water bottle, doesn't leak
Color: Camo Cool, Size: 24 Ounces
Headline: Finally, a Water Bottle That Lives Up to the Hype!⭐⭐⭐⭐⭐"I’ve tried almost every popular water bottle brand out there, and the Owala FreeSip is genuinely in a league of its own. The 2-in-1 spout is a game-changer—I love being able to sip upright through the straw or tilt it back to chug. It makes drinking water so much easier.It’s completely leakproof when locked, which I’ve tested in my gym bag multiple times. Plus, the cover keeps the mouthpiece sanitary. It keeps my water ice-cold for over 24 hours, even sitting in a hot car. And let's talk about the colors—they are so fun and vibrant. I got the 24oz, and it fits perfectly in my cup holder. If you’re on the fence, just buy it!"
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Reviewed in the United States on May 4, 2026
★★★★★ 4
Great Water Bottle but failed after one year.
Color: Foggy Tide, Size: 32 Ounces
I purchased this Owala water bottle a little over a year ago and it was in my top favorites. The flip-up lid is a great feature, and I like that you can use it with or without the straw. It’s completely leak-proof, comfortable to carry, and has held up well with daily use. My only complaint had been that it doesn’t fit in most standard cup holders.
But after about a year, I did notice that it stopped keeping my water cold as long as it used to, and the ice began melting much faster. I reached out to Owala customer service but unfortunately never received a response.
Even with that issue, I still prefer the lid design over brands like Yeti or RTIC, so I would consider purchasing another Owala in the future. If you value a convenient, leak-proof lid design, it’s definitely worth considering. I just sincerely hope the insulation lasts longer on the next one.
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Reviewed in the United States on May 18, 2026
★★★★★ 5
The BEST water bottle on the market!
Color: Denim, Size: 32 Ounces
I am in love with this water bottle! I take it everywhere with me, and I love that it has a built-in straw. The color is perfect as well. It motivates me to get my water intake in, and it helps me meet my fitness goals. It is easily washed in the dishwasher, and I don't have to worry about it breaking. It keeps the temperature of my water perfect- whether I add ice or not. It is sturdy but not too heavy.
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Reviewed in the United States on May 26, 2026
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