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Description
Rat TGF-b2 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: 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: 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. 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. 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. 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. 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. 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 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 1000pg/mL). Then dilute to the following concentrations: 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, 15.625pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 1000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 500pg/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 a Transforming Growth Factor Beta 2 (TGF-b2) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Transforming Growth Factor Beta 2 (TGF-b2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Transforming Growth Factor Beta 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Transforming growth factor β2 (TGF-β2), also known as glioblastoma-derived T-cell suppressor factor (G-TSF or BSC-1), is a secreted protein called a cytokine. It performs many cellular functions and plays a crucial role in embryonic development. It is an extracellular glycosylated protein and is known to inhibit the action of interleukin-dependent T-cell tumors. It regulates various processes, including angiogenesis and cardiac development. Its immune-regulating effects include: reducing inflammation: Through regulatory T cells, it helps balance the immune system and reduce inflammatory substances; reducing allergic reactions: It reduces the level of allergies caused by IgE (immunoglobulin E), reducing hypersensitivity; improving food tolerance: It helps build tolerance and mitigates the risk of allergies to foreign proteins in food; and protecting against viral and bacterial invasion: It promotes IgA (immunoglobulin A) secretion, which helps the intestinal mucosa develop and resists foreign invasion; and protecting against influenza A virus (H1N1) infection, reducing the severity and duration of the disease. | |||||||||||||||||||||||||||||||||
| 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 | 15.62-1000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.1 ★★★★★
Based on 375 reviews
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Product Reviews
★★★★★ 5
AMAZING debut novel!!!
Format: Kindle
Plot ⭐️⭐️⭐️⭐️
Spice 🌶️🌶️.5
Romance 💘💘💘
Vibes ⭐️⭐️⭐️⭐️⭐️
Dual 1st person POV - Ara (26) & Rogue (39 - but looks mid-20s: they can live hundreds of years so this isn't that large of a gap as it could've been which I heavily appreciate lol)
Tropes: enemies to lovers, fae/human wars (deep hatred for each other), shifters (dragons- MMC can only partial shift with wings), one horse, one bed, touch her and d!e, found family, abduction turned to freedom
The Last Storm is the debut novel from JD Linton and let me tell you, you guys NEED to read this. The plot was engaging and the editing was was amazing (especially for a debut novel). Our FMC, Ara, is stuck in her gilded cage longing for a life outside of her small town. She uses her books to escape and live vicariously through the pages (honestly, relatable). After her father announces her betrothal to her childhood friend (to whom she has no romantic feelings for), Ara tumbles unknowingly into a desperate plot trying to stop the humans from slaughtering the Fae.
As one can expect from an enemies to lovers / kidnapper/captive romance, Ara fights her attraction and lust towards our MMC, Rogue (the King of the Fae), for as long as she can. Upon seeing Ara for the first time, Rogue is instantly aware that she is his fated mate (not a spoiler). Since she is the General's only daughter, he plans to abduct her and use her as leverage to stop the brutality. During Ara's time in Rogue's captivity, their banter and chemistry continue to rise until they finally boil over and come together (quite literally, and many times I may add 😉).
Here's what I LOVED:
- Rogue continuously seeks advice from his elders and deeply respects their opinions and life experience and tries to implement their recommendations
- Rogue makes many mistakes in the beginning but we see him actively work on not repeating them as the book progresses. The level of self-awareness and his ability to change his behavior was impressive
- The magic system is intricate and we have only scraped the surface. As the series continues and Ara progresses in her powers, I'm sure we'll get to see more of this. I absolutely LOVE the messaging system that is used in this book.
- Ara's struggles are so human and so raw. She is experiencing so much guilt and pain and hurt and getting to see her work through each of these emotions is inspiring. Especially as her and Rogue get closer and she learns she can lean on him as well, that she is not alone.
- While this is the start of a series, there is NO cliffhanger! There's a bit of a teaser of something major that is going to happen at the start of the next book, but it's not a cliffhanger in the sense that we aren't sure if someone is going to live or d!e or if they'll be separated. For that, I am very thankful! This book was so much fun that I will definitely be returning to book 2, even if it takes several months (or longer since this is an debut author) to publish!
- Lastly, the cover is GORGEOUS! And I love the title!
I'll copy a few of my favorite quotes below so you can have a little taste of the author's writing and the world she's cultivated. 😊
Top Highlights from The Last Storm
On days like this, when my heart was heavy and my mind clouded, I resorted to books— to escape, to forget, to find freedom where I had none.
If I were to marry him, my face would always be turned to the window, searching for more, and if not that, I would be a shell of the person I am now.
I stepped back to admire her, thr0bbing at the sight. She was the most beautiful woman I had ever seen. To ever exist. Nothing, no one, had ever deserved to be worshiped more. All men should be made to kneel before her. But she would have to settle for me.
The taste of her met my t0ngue as my scent merged with hers, forever branding her. Mine. I l!cked the wound. Hers. Completely and utterly hers.
I didn’t claim her in ownership. I claimed her as my one. Devoted myself to one. With that mark, my body and soul were bound to her. I would never be with anyone else, emotionally or physically. It would be her or no one, until my last breath.
“Scream my name. Let everyone know who I belong to.”
I had never really cared about the weather before, but now, clear skies meant everything to me, and I was grateful to see another calm morning.
“There will never be another woman for me.” He paused. “Ever.” I stilled at his words. “What… Why?” “This”— his thumb slid down across the mark—“ is a symbol of… surrender. I know you believe that it was my claim upon you, but it wasn’t. It never was. I bound my body and soul to you, little storm.”
“I also know that it is more than this tiny, insignificant mark on your skin that binds me to you. It’s you. All of you. Your strength and resilience. Your determination to endure no matter what fate throws at you. Your love for love and stories and hope. You are entirely the opposite of everything that I am and I would gladly wear your shackles if it meant I could have you.”
My mate. Mine. And then everything shifted and I understood. I understood everything. The surrender. The deep, soul-craving longing. Bound. I was bound to him. Body and soul. Entirely his. “I would’ve waited forever,” he whispered back, understanding.
Seriously, everyone.. add this to your TBR!!
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Reviewed in the United States on November 14, 2022
★★★★★ 4
A Decent Fae Romance
Format: Kindle
** 3.5 Stars rounded up to 4 **
First off I want to say that I found the author, J. D. Linton, on TikTok right before the release of the sequel. I bought the first 2 books to support her release, so congrats to her for the release of the sequel!
Overall, I liked the story & plot, I liked the characters, and I liked the spice. The downfalls: not enough development between the characters for the romance, and it leaned a little heavy into the tropes. A lot of stuff is told to you, which could have been used as devices to drive the plot and emotions of the characters.
🛑🛑 Mild Spoilers ahead, I tried to hide the obvious ones 🛑🛑
This is very obviously a fated mates, which is a trope I usually really enjoy. But I wish the relationship between Ara & Rogue had much more development before it's realized that they're mates, instead of Rogue knowing almost immediately and using it for nefarious purposes (at least in the beginning before they get to know each other). By revealing this within the first quarter of the book, I feel like it leaves less room for them to fall for each other organically (albeit with help from the mating bond) and they love each other because of the mating bond. I was disappointed as soon as Rogue know (literally only 5% in) and I literally made a note: "As much as II love a good mating trope, I wish we had to work for it a little more. Where's the fun in just telling us?" I believe that by holding out and feeding the reader snippets of a potential bond, it would've been more rewarding as a reader.
I also wish there was more world building - we are told of a war between human & fae but don't get to really experience any of it. Ara is sheltered in her human home, then sheltered in Rogue's castle. There's bits and pieces about what the war has done on either side - but we're more told of the aftermath and don't really experience any of it. Ara's father is supposed to be the king's #1 general - yet he is at home with his family & with Ara for the first couple chapters. Her, her family, nor her village seem to be affected by the 10 year war going on on their borders. I wish there was a little more setup to make this conflict - an actual war - feel more than a skirmish between fighting territories.
Linton could also be a little repetitive - with the biggest culprit being when Ara is upset she "brings [her] knees to [her] chest]" and either sits like that or cries. Every time she is upset this phrase is mentioned. I would get it if this was her crutch, or how she copes with grief and stress, but that should be explained why she does it so often or it becomes repetitive. I started to get annoyed with how often she would sit like this solely because it happens every couple chapters.
However, I did really like the spice. I love an enemies to lovers trope, especially when it results in spicy scenes. The spicy scenes weren't anything new, but they were fun. Wish there were more but that could also just be me - there is no such thing as too much spice 😂
Would I read again? Probably not, I'm super picky with rereads. Although I did genuinely enjoy my first read through!
Will I continue the series? Probably, at least for the sequel. As for #3, kind of depends on where I am with my TBR once that is released
All in all an enjoyable, fast paced read
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Reviewed in the United States on March 8, 2024
★★★★★ 3
I love indie authors
Format: Kindle
Let me preface this by saying—writing a book is HARD! Coming up with characters so real that they take on a life of their own, building an entire world, the political/magic system. Designing all of this is no easy feat.
That being said.
This book had so. Much. Potential. I was so excited to read this book, and I plan to finish it for the sake of finishing it. But. At this point, I would have set I’d aside as a DNF. The book could have benefited from some form of a developmental editor, or an in depth beta reader. I will say this. Within 5 chapters, there are so many… phrases that I’ve highlight that I’ve latched onto. Phrases about books and storms that were written BEAUTIFULLY! So, bravo Linton for hitting the nail on the head as to why readers disappear like they do!
However, 5 chapters in and I can already guess where a majority of the story is going. But that’s also because I read like a mad-woman and have read this particular type of story, many different ways. Enemies to lovers where the FMC isn’t who she thinks she is. I am all about supporting indie authors. BUT. I also feel like criticism should be constructive, and not degrading. So if I could give this book a 4 1/2, strictly because I know the work the author put into this, I would. So if you’re looking for an easy read with characters that aren’t hard to follow, look no further! They are easy to love and easy to care for. One of the biggest issues they lacked, to me, was depth and plausible reactions to their situations. JD, you have done BEAUTIFULLY writing this book. I applaud and will continue to buy your books in the future. My BIGGEST recommendation is to definitely hire some form of an editor for any upcoming books. Or in turn, I will be happy to beta read for you. Should my opinion change of the book by the time I finish, I will happily get on here and say I was wrong, delete this review and post a different one. Until then…
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Reviewed in the United States on November 20, 2022
★★★★★ 5
this book set my soul ablaze! <3
Format: Paperback, Format: Paperback
"i had never really cared about the weather before, but now, clear skies meant everything to me, and i was grateful to see another calm morning."
this book. this book!
i loved the last storm so much. the writing style. the descriptions. the world-building. the characters. the plot twists. the tropes. the sexual tension. the—everything. everything was magic.
the last storm follows our two main characters, ara and rogue, giving us dual POV from both characters (which i loved, btw). ara, a human girl who has been locked away in her father’s estate most of her life, just wants to see the world. all she dreams of is seeing what else is out there. but when her father announces her engagement, she knows that dream will become nothing more than just that—a dream.
rogue, the fae king, is tired of the attacks being rained down on his people. in hopes of finding out the human king adon’s secrets, rogue infiltrates auryna’s borders. in his last resort to gain information, he visits the local pub. to his surprise, the general’s precious only child is sitting at the bar, drink round after round of mead. now he just needs to figure out how to take her without anyone noticing.
first and foremost, let’s talk about the endless list of my favorite tropes and aspects that this book had.
›› enemies to lovers
›› fated mates
›› one bed
›› the chosen one
›› elemental magic
›› actually good and shocking plot twists!!!
›› badass female lead
›› morally-grey love interest
›› fae/human war
›› force proximity
›› touch her and die
›› who did this to you?
›› captor/captive
›› praise k!nk (panting profusely)
“you are entirely the opposite of everything that i am, and i would gladly wear your shackles if it meant i could have you.”
it’s been a long while since i read a book i liked this much. but i just loved this book. it set my soul ablaze. thank you to the author for writing this beautiful story and for blessing me with an eARC! i loved it so much that i immediately bought the paperback upon release!
every aspect of this book was just beautiful. i was blown away by the way the world was described, the way feelings were portrayed, the way the elements were used in the fae’s magic. it just—AHHH! i just absolutely adored it all. i cannot wait for the second book to release next year!
also the way he calls her “little storm” sets my heart on fire.
this was a fast-paced read and if you are a lover of acotar, fbaa, deal with the elf king, or any other similar books, then please stop everything you’re doing and read this book right now. you won’t regret it.
thank you again, jd linton, for giving me the privilege of reading your arc and for blessing this world with the world you created. <3
"something about him pulled me in, like a moth to a flame, and it felt as if i was just waiting for the inevitable burn that came with flying too close to the fire."
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Reviewed in the United States on November 15, 2022
★★★★★ 4
Fun, both heartwarming and heartbreaking
Format: Kindle
Only my second first person written selection, I am still getting used to that aspect, but unlike my first, I enjoyed that the story was told through both MCs. A great enemies to lovers, forced proximity, fated love etc, that resonated to me. There were some small twists that I could see coming, but also a few that I didn’t quite see until the characters were also seeing. Personally, I am more interested in the story than the spice, but with that said, it was well seasoned! I am kind of new to the spice world so I can’t say for sure how this would rate, but it definitely had some heat. I am very glad I happened across this author, and I do plan on also reading the next book….if nothing else, just to see for myself the “transformation” of the characters I’ve grown to love!
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Reviewed in the United States on May 23, 2024