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Description
Rat NT-ProBNP 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. Preparation of standard gradient working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 5000pg/mL). Then dilute to the following concentrations: 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312.5pg/mL, 156.25pg/mL, 78.125pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 5000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 2500pg/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 enzyme conjugate are sequentially added to microwells pre-coated with an N-Terminal Pro Brain Natriuretic Peptide (NT-ProBNP) 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 N-Terminal Pro Brain Natriuretic Peptide (NT-ProBNP) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat N-Terminal Pro Brain Natriuretic Peptide ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | N-terminal pro-brain natriuretic peptide (NT-ProBNP or BNPT) is a 76-amino acid N-terminal inactive protein precursor, from which brain natriuretic peptide (BNP) is cleaved. Blood levels of both BNP and NT-ProBNP are used to screen for and diagnose acute congestive heart failure (CHF) and to determine the prognosis of heart failure, as both markers are often elevated in patients with a poor prognosis. Plasma concentrations are also often elevated in patients with asymptomatic or symptomatic left ventricular dysfunction and are associated with coronary artery disease and myocardial ischemia. | |||||||||||||||||||||||||||||||||
| 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 | 78.12-5000pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.7 ★★★★★
Based on 27 reviews
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Product Reviews
★★★★★ 5
Poems worth analyzing
Format: Hardcover
Each chapter is a close reading of a poem, insightful and jargon free. Maybe a bit more context and biography for my taste, but still helpful. Reawakens the reading of poetry.
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Reviewed in the United States on February 27, 2026
★★★★★ 5
I once thought I knew about poetry. My eyes and heart have now been opened.
Format: Hardcover
This is the first time I have read any of her essays. Magnificent! Such insight in how each poem is constructed and what she feels about each poem. Reading this work has helped me and enriched my poetry reading by showing me new depths found in each poem. If you love poetry, give yourself a treat. Read this work.
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Reviewed in the United States on October 6, 2025
★★★★★ 5
Beautiful and Hopeful...
Format: Kindle
“Olivia was right. The point of the choose your own adventure books was just that: Choice. It wasn’t about where you ended up; it was about the decisions you made to get there. And I don’t want to skip to the end anymore…” – Rosaline, When You Were Mine
If I could give this book 500 hearts I would, but my rating system only goes up to 5.
I’ve read a lot of great books lately, but When You Were Mine by Rebecca Serle surpasses 95 percent of them. This book was amazing from start to finish. I am a lover of all things Shakespeare, so when I saw the synopsis for this book it was an automatic must read, plus guys the cover is so pretty!
I absolutely loved the format of this book and the writing was exceptional, but I think what I loved the most were the characters. Serle did an amazing job of shaping the characters in this book, from Rosaline, Charlie and Olivia to Ben Rob, Juliet and of course Len, who by the way I loved from the second he was introduced.
The plot and pacing of the book were perfect. I didn’t feel rushed when reading about Rosaline and Rob or Rosaline and Len. Seeing things unfold throughout the story and finding out what happened to cause the two sets of Caplet’s to drift apart was a bit twist. I wasn’t exactly that at all and the fact that Serle was able to surprise me at countless turns throughout the book sort of makes me want to glomp her with hugs, because that’s hard to do to me.
There were so many amazing parts in this book that it’s difficult to choose favorites, but at the same time I had such strong emotions to certain parts of this story that I feel I need to mention them. First the Fall Back Dance. My god…I can’t even tell you how a part of my heart genuinely broke for Rosaline. I felt her pain, like literally. I got all teary eyed and my chest tightened up and I think I was like ‘OMG Rebecca you’re so mean!’ Haha. But the scene was so well written, that I experienced what happened there with her.
Another scene that touched me was the piano scene with Len. It will probably go down in my like top ten scenes in a book between two characters ever. The way Serle describes what’s happening and the emotion flowing through both characters and Rosaline’s reaction to the music, just, I can’t even. I wouldn’t use gifs to explain my emotions right now, but I’m not sure any of them would accurately capture what this books has done to me.
The third scene that really broke me and my raw hatred of Juliet was the kitchen scene between her and Rosaline. All through the first couple of acts of the story I was hardcore hating on Juliet. Like reach into the book and pop her on the head kind of hate. But after this scene…I just couldn’t do it anymore. Serle managed to take the essential villain of the story and feel empathy for her.
God, this entire book was just made of so much win. I want to draw pink fluffy hearts around it and love it forever. I am so glad I bought the hardcopy version instead of the Kindle version because I will definitely be reading this again…and then most likely again. I’m currently writing this while I shove the book at my sister and tell her to read it so she can love it as much as I do.
If you haven’t read When You Were Mine, for the love of god get on it. This has been on my reading least for so long and I’m just getting around to it now, but honestly I wish I would have pushed it up on my list. Amazing, no other words for it, definitely check it out if you get the chance and if it’s sitting on your list, well, move it up it’s worth it!
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Reviewed in the United States on March 20, 2014
★★★★★ 4
Refreshing Take On a Classic!
Format: Kindle
I have always been a sucker for "fractured fairy tales" and classic stories being retold by new voices (I think that's why I love Disney movies so much!). I have also always been a sucker for Shakespeare, in awe of the way he uses the dynamics of relationships to create stories that are as mind-blowing today as they were centuries ago when they were written. So when I heard about Rebecca Serle's When You Were Mine, it was essentially book candy on which I wanted to overdose!
Set in modern day southern California, When You Were Mine is the retelling of William Shakespeare's Romeo & Juliet from the perspective of Rosaline (are you as absolutely giddy over the premise as I was?! It's probably sad that I'm not being sarcastic at all right now!). If you don't remember Rosaline from middle school English class, she's the girl Romeo was madly in love with at the beginning of the play; until he met Juliet, of course. The first thing I thought of when I heard about this book was, "Why hasn't anyone else done this already?? Why didn't I think of this?!" (Honestly, maybe it has been done, but not to my knowledge).
So the idea is great, but what about the execution? I had high hopes when I started reading and overall I feel like the story delivered, but I admit I had to keep reminding myself that this is a contemporary Young Adult novel. Rob Montag and Rose Caplet are high school students who have been next door neighbors most of their lives. They have grown up as best friends but their friendship is beginning to blossom into more, much to Rose's excitement. But when her cousin Juliet moves back to town (after years of being estranged from Rose's family for a political scandal, no less), Rob becomes more interested in Juliet, leaving Rose (friendship and all) in the dust.
The story line and narration was very "high school" and I found myself becoming exhausted at times, like I was truly listening to a teenage girl babble. In one sense, this made me consider Serle as an incredible writer because she was authentically able to capture the voice of teenage Rose and maintain that voice throughout the novel. However, it did make reading taxing at times; almost everything Rose said consisted of her reflecting on something that had happened to her and her friends previously (think, "One time. At band camp...."). This did not slow my reading down necessarily (I inhaled the novel in 1 day), but all of the side stories definitely affected the pace of the novel. I think it also made me feel self-conscious, like, is this what I sounded like at this age? (Don't answer that, Mom!).
Aside from this Young Adult novel sounding like a super young adult (trying to emphasize that I can't really fault the novel for my biggest complaint), I really enjoyed the characters, the plot and especially the parallels between Shakespeare's version. The main characters were likable, especially Rose and her best friends. The only characters I wish the story would have given more dimension to are Rob and Juliet. I thought Rob was well developed until he fell for Juliet; I couldn't understand why such a great guy would go for this snotty girl, which made me feel like maybe I didn't understand Rob at all. (Of course, I'm sure Rose felt the same way and she is the narrator, after all). I was disappointed that there was not more depth to Juliet as well, but I suppose it's only fair that her side was mostly omitted from this story, as Rosaline's was cut out of the original, right?
What makes this novel stand out to me is how many emotions are vicariously experienced while reading: hopeful, happy, disappointed, angry, jealous, frustrated, sad, guilty, lonely, desperate, grieved, peaceful, and right back around to hopeful (there are probably a dozen more I left out). It's an extreme love story (we all know how it ends), but told by the perspective of Rose is refreshing and introduces the point of view of those who lived after Romeo and Juliet died. What is most rewarding toward the end is how Serle paints the picture of Rose's grief on top of a shattered heart. Without giving too much away, I feel like the conclusion was heartwarming and hopeful, despite its tragedy. Serle gives us something familiar that we can mourn differently than the first time we heard this tale.
Bottom Line: If you're a sucker for Romeo & Juliet, this is a must-read. If you're a sucker for YA I highly recommend this, too! If your reading preferences are a little more stuffy, then you might not be impressed (but that's your loss!).
[...]
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Reviewed in the United States on July 13, 2012
★★★★★ 3
Teenage love story
Format: Paperback
Not my cup of tea, takes place in High school so I think it was just a little young for me, couldn't get into the drama. Love her other books though!
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Reviewed in the United States on August 15, 2023