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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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4.9 ★★★★★
Based on 2347 reviews
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Product Reviews
★★★★★ 4
Well…
Format: Kindle
I enjoyed the book, it had an interesting plot and magic system. The book is full of smirks, hissing, and crying. OMG the crying! The FMC cries from front to back of the book! I absolutely loved Daggermouth and read this book bc that, but I feel like she had a different editor maybe?! Idk what happened, but this book would have been so much better without the MMC smirking on every page that the FMC was crying on.
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Reviewed in the United States on May 29, 2026
★★★★★ 5
Top Book of 2024
Format: Kindle
May Contain Spoilers!
Where do I even begin?
I'm still in shock of how amazing this book was, how it touched my heart, and now how I have to wait for H. M. Wolfe to write the second. Because those last few chapters...went from 0 to 100 quick. So much new information, new relationships and loss, that now the tides have turned, and we NEED more.
This is a fantasy book of love, loss, the power of friendship, and finding one owns strength. TBOC is about Hyacinth whose home is at Asrai’s Academy for Orphans. She was torn between the safety of home or leaving it all behind for a chance at a new life. But when a mysterious messenger appears, whispering rumors of a God among Fae, everything Hyacinth thought she knew about the Realms comes crashing down, and she must now make a new journey.
This book ate me up and spit me back out. I was not anticipating the emotions I would feel within these pages. I'm talking I was laughing, then I was swooning, then I was on the edge of my seat, then I'm ready to slap someone, then I'm rooting so hard for certain characters I don't wan to stop, then my heart is broken, then I'm excited af.... it was all an emotional rollercoaster. The last few chapters contained a lot of new information, so of course as much as it doesn't leave off on a complete cliffhanger, I was over here like scraping the bottom of an ice cream cup, wanting more.
The magic system starts out ambiguous, but as you continue reading you start to get more and more to build off of and start to piece together the world H. M. Wolfe created. TBOC brings in eight main characters that test alliances, grow in friendships, fall in love, become a team, and even break bonds.
The friendship among the girls was everything. Ata is a spit fire, Pria is sweet yet terrifying. The banter and protectiveness these three had for one another made me want to join in with them for breakfast. I liked how there was multiple romances going on at once. Yes we have the main characters, but it just wasn't always about them, and it was great to see how others shifted and grew too.
And last the romance, I've never rooted so hard for two characters. We're talking slow burn between them. In the beginning you're not sure how this is all going to play out, but Landers is a tatted, "touch her and die" kind of MMC, which who cannot love that? Hyacinth definitely had to grow within the chapters and learn how to be strong, because sometimes she was making me mad and I just wanted to step in there for her. But Landers was everything she needed and it was so well written, which is why by time you get to the end you only want more of them!
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Reviewed in the United States on September 24, 2024
★★★★★ 3
cliffhanger, no 2nd book in sight
Format: Kindle
The book itself was pretty good. The plot is unique and full of suspense and action, although some of it felt redundant. I particularly enjoyed the realistic growth cycle of the female lead from abuse survivor with low self-esteem to badass warrior.
I never would have read it though if I had known this is not complete, and there is no information available on the next book.
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Reviewed in the United States on November 17, 2024
★★★★★ 4
4.5
Format: Kindle
4.5
The Book of Cin relies heavily on the progression and growth of our fmc, which I don't hate. It shows us the girl she was and what she allowed and who she then grows into by the end due to all the events that happens within the book. This journey - physically, mentally, and especially emotionally - takes us through her current and past experiences and expresses how one can grow from trauma. Her support group is invaluable, and the new relationships she forms prove to be just as prominent to her and her growth. This is truly a story that thrives on the characters growth throughout the book and sets us up in how to see her for the coming books.
My half star comes from the pacing. Though I *thoroughly* enjoyed the ending and seeing this growth take full effect, I believe that it came with just too many pages. Sitting at about 500, we get intimate and detailed scenes that we could have maybe gone without. Oftentimes, I found myself daydreaming away from the book until something more prominent peaked my interest again. I understand the depth the book goes into to fully establish this character development and the setup that this provides for the rest of the series, but I truly do believe there were scenes that could have been cut out to help with the pacing.
If you're not into slow pacing (and usually I'm not), this book is still worth the read. The ending more than makes up for it, and it is absolutely detrimental to the story that we get the events that occur and see the development take shape in front of our eyes. It makes you fist pump the sky by the end.
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Reviewed in the United States on May 20, 2025
★★★★★ 5
All the stars! Emotional! Incredible!
Format: Kindle
𝘔𝘺 𝘩𝘦𝘢𝘳𝘵 𝘥𝘪𝘥𝘯’𝘵 𝘫𝘶𝘴𝘵 𝘴𝘵𝘰𝘱 𝘣𝘦𝘢𝘵𝘪𝘯𝘨—𝘪𝘵 𝘴𝘩𝘢𝘵𝘵𝘦𝘳𝘦𝘥 𝘪𝘯𝘵𝘰 𝘢 𝘮𝘺𝘳𝘪𝘢𝘥 𝘰𝘧 𝘧𝘳𝘢𝘨𝘮𝘦𝘯𝘵𝘴 𝘵𝘰 𝘵𝘩𝘦 𝘥𝘢𝘳𝘬𝘦𝘴𝘵 𝘤𝘰𝘳𝘯𝘦𝘳𝘴 𝘰𝘧 𝘵𝘩𝘦 𝘷𝘰𝘪𝘥 𝘤𝘰𝘯𝘴𝘶𝘮𝘪𝘯𝘨 𝘮𝘦.
This was one of those books I have a hard time reviewing because I struggle to find the words to adequately express how incredible it was! This book deserves every single star! It is heart-wrenching, emotional, and heavy. The depictions of self-worth, guilt, loss, love, and everything in between were done beautifully. There are multiple romances to root for and the development of the plot/world was executed brilliantly. It has an amazing set of characters that will bring out not only all the rage/hate but also sacrifice that will impact your soul! The ending of chapter 19 will live rent free in my mind and is now one of my top favorite moments from any book and why Landers will forever take a top spot in my all-time favorite book boyfriends! The depiction of what true friendship should be is also well represented! Ardan stole my heart as the best side character and if you’ve read this we need to talk (iykyk)!! I absolutely need the next book to see what happens next!!
“𝘐 𝘴𝘩𝘰𝘶𝘭𝘥𝘦𝘳𝘦𝘥 𝘺𝘰𝘶𝘳 𝘱𝘢𝘪𝘯, 𝘯𝘰𝘵 𝘢𝘴 𝘢 𝘵𝘦𝘴𝘵𝘢𝘮𝘦𝘯𝘵 𝘵𝘰 𝘺𝘰𝘶𝘳 𝘸𝘦𝘢𝘬𝘯𝘦𝘴𝘴, 𝘣𝘶𝘵 𝘣𝘦𝘤𝘢𝘶𝘴𝘦 𝘮𝘺 𝘰𝘸𝘯 𝘴𝘵𝘳𝘦𝘯𝘨𝘵𝘩 𝘤𝘳𝘶𝘮𝘣𝘭𝘦𝘥 𝘪𝘯 𝘵𝘩𝘦 𝘧𝘢𝘤𝘦 𝘰𝘧 𝘺𝘰𝘶𝘳 𝘴𝘶𝘧𝘧𝘦𝘳𝘪𝘯𝘨. 𝘐 𝘤𝘰𝘶𝘭𝘥 𝘯𝘰𝘵 𝘴𝘵𝘢𝘯𝘥 𝘪𝘥𝘭𝘦 𝘸𝘩𝘪𝘭𝘦 𝘐 𝘭𝘪𝘴𝘵𝘦𝘯𝘦𝘥 𝘵𝘰 𝘴𝘤𝘳𝘦𝘢𝘮𝘴 𝘰𝘧 𝘢𝘨𝘰𝘯𝘺 𝘤𝘰𝘮𝘦 𝘧𝘳𝘰𝘮 𝘢𝘯𝘰𝘵𝘩𝘦𝘳 𝘸𝘰𝘮𝘢𝘯 𝘐 𝘤𝘢𝘳𝘦 𝘧𝘰𝘳. 𝘐𝘵 𝘸𝘢𝘴 𝘯𝘰𝘵 𝘮𝘺 𝘱𝘭𝘢𝘤𝘦 𝘵𝘰 𝘵𝘢𝘬𝘦 𝘵𝘩𝘢𝘵 𝘧𝘳𝘰𝘮 𝘺𝘰𝘶, 𝘺𝘦𝘵 𝘐 𝘸𝘰𝘶𝘭𝘥 𝘥𝘰 𝘪𝘵 𝘢𝘨𝘢𝘪𝘯, 𝘧𝘰𝘳 𝘢 𝘵𝘩𝘰𝘶𝘴𝘢𝘯𝘥 𝘮𝘰𝘳𝘦 𝘩𝘰𝘶𝘳𝘴, 𝘪𝘧 𝘵𝘩𝘢𝘵 𝘮𝘦𝘢𝘯𝘵 𝘺𝘰𝘶 𝘥𝘪𝘥 𝘯𝘰𝘵 𝘩𝘢𝘷𝘦 𝘵𝘰 𝘴𝘶𝘧𝘧𝘦𝘳 𝘦𝘷𝘦𝘯 𝘢 𝘴𝘦𝘤𝘰𝘯𝘥 𝘭𝘰𝘯𝘨𝘦𝘳.”
“𝘐 𝘢𝘮 𝘯𝘰𝘵 𝘩𝘪𝘮. 𝘐 𝘸𝘢𝘯𝘵 𝘵𝘩𝘦 𝘴𝘩𝘦𝘭𝘭 𝘰𝘧 𝘺𝘰𝘶. 𝘐 𝘸𝘢𝘯𝘵 𝘵𝘩𝘦 𝘣𝘳𝘰𝘬𝘦𝘯, 𝘴𝘤𝘢𝘵𝘵𝘦𝘳𝘦𝘥 𝘴𝘩𝘢𝘳𝘥𝘴. 𝘐 𝘸𝘢𝘯𝘵 𝘵𝘩𝘦 𝘵𝘦𝘢𝘳𝘴, 𝘵𝘩𝘦 𝘯𝘪𝘨𝘩𝘵𝘮𝘢𝘳𝘦𝘴, 𝘵𝘩𝘦 𝘱𝘢𝘪𝘯. 𝘐 𝘸𝘢𝘯𝘵 𝘺𝘰𝘶. 𝘈𝘭𝘭 𝘰𝘧 𝘺𝘰𝘶. 𝘛𝘩𝘦 𝘧𝘦𝘢𝘳, 𝘢𝘯𝘥 𝘦𝘷𝘦𝘳𝘺 𝘧𝘳𝘢𝘨𝘮𝘦𝘯𝘵𝘦𝘥 𝘱𝘪𝘦𝘤𝘦 𝘰𝘧 𝘺𝘰𝘶𝘳 𝘣𝘭𝘢𝘤𝘬𝘦𝘯𝘦𝘥 𝘴𝘰𝘶𝘭; 𝘐 𝘸𝘢𝘯𝘵 𝘵𝘩𝘰𝘴𝘦 𝘱𝘢𝘳𝘵𝘴 𝘵𝘰𝘰.”
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Reviewed in the United States on June 23, 2025
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