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Description
Mouse 5-HT3A 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 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/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 conjugate are sequentially added to microwells pre-coated with a 5-Hydroxytryptamine Receptor 3A (5-HT3A) 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 5-Hydroxytryptamine Receptor 3A (5-HT3A) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse 5-Hydroxytryptamine Receptor 3A ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The serotonin receptor 3A (5-HT3A) is a protein encoded by the HTR3A gene. The gene product belongs to the ligand-gated ion channel receptor superfamily. This gene encodes subunit A of the type 3 receptor for serotonin (5-HT), a bioactive hormone that functions as a neurotransmitter, hormone, and mitogen. Activation of this receptor induces a rapid depolarization response in neurons. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.8 ★★★★★
Based on 1161 reviews
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Product Reviews
★★★★★ 5
An action-packed dark romantasy
Format: Kindle
I loved this book! Queen of Roses is an Arthurian-inspired dark romantasy that is the first book in the Blood of Fae series. The story follows Morgan, the princess of Camelot who is rumored to be part fae. Fueled by prejudiced hatred and a mistrust of fae blood, Morgan’s abusive father strips her of her birthright and hands it to her half-brother, Arthur. Instead of becoming queen, Morgan is commanded to join the temple of the goddesses when she comes of age.
However, Arthur turns into a psychopathic, power-hungry, fae-hating king as he ages. He develops malevolent plans and commands Morgan to find an ancient weapon with legendary power. Although Morgan is wary of Arthur’s intentions, she embraces the opportunity to go on a journey and potentially change her fate.
The story picks up from there and we follow Morgan on her quest to find the ancient relic. It’s full of high stakes adventure, mystery, tension, banter, forced proximity, hidden magic, self discovery, and betrayal. This first installment of the series intricately develops the world building and character development. There’s little romance in this book, but it is evident that it is a slow burn that will continue to develop throughout the remainder of the series.
Overall, I loved the world building, the epic fantasy, Morgan’s journey of self discovery, and all of the twists and turns that set the stage for the future installments. I can’t wait to see what happens next!
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Reviewed in the United States on April 7, 2024
★★★★★ 4
Arthurian Fae Quest…say less.
Format: Kindle
A fae centered Arthurian tale unlike any I’ve read so far. The author did a great job at descriptive world building, with scenes easily playing out in my minds eye. There was plenty of action, suspense, and even a touch of horror. An enemies to lovers, slow burn romance, a quest, with plot twist and turns aplenty. There was a love triangle, which I’m not usually a fan of but, it played out well in this story line. The FMC, Morgan Pendragon, was so blatantly naïve, yet I typically expect as much in a ‘book one’ of a series, especially one that features a fairly sheltered princess. I was happy to read that in spite of this, she still showed a strong sense of morals, fire, and spine. Now our MMC? Kairos Draven, aka Void’s Edge. Oh, how I’m a sucker for a smoking’ hot grumpy warrior alpha with a witty mouth, and a strong sense of “touch her and die” attitude, so you know who held all my cards. That ending? Just made me swoon all the harder. Now add a battlecat that rivals the size of a horse…and well Ms. Briar Boleyn you have well and truly stolen my heart. I’m excited to see where the story goes from here, and follow along to see more of the characters growth. I went into this story fairly blind, and I think I enjoyed it all the more because of it.
Once the story got going, it had me in an absolute chokehold and it was difficult to put down.
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Reviewed in the United States on May 12, 2024
★★★★★ 3
Not a bad start
Format: Kindle
3 stars
Thank you Netgalley and Briar Boleyn for the ARC!
A camelot/king Arthur retelling with fae. I was hooked by the idea of this book immediately and was eager to jump into this world.
• slow burn
• enemies to lovers
• who did this to you
Morgan Pendragon watched her mother die by her father's hand when she was just eight years old, hiding under the bed. Morgan is believed to have the tainted blood of the fae in her veins and is cast aside so that her fathers illegitimate son, Arthur, can become the king. She's seen his cruel treatment of the fae firsthand, so when he sends her on a journey to find a fae weapon she seizes the opportunity to do more with her life. Along the way, she finds more than she could have imagined.
I don't know a whole lot about King Arthur and Camelot but I had a lot of fun with this story! The plot has some similar tropes to popular romantasy books (From blood and ash) but there's enough originality here that it doesn't feel like I'm reading a copy. I liked how the fae were different in appearance than what is typical in most fantasy books I've read. In this book they have blue hair, violet skin and a wide range of other characteristics. I thought that the world building was easy to follow and I could easily immerse myself into this world. After reading the blurb I kept wondering when she was going to go on the journey to find Excalibur and it doesn't happen until around the 45% mark.
The story is a bit slow at times but starts to pick up once they begin their journey to find Excalibur. The John Wick style Inn was a fun concept that I enjoyed reading about.
There are a lot of similarities to this and FBAA and I would have liked to have it be a little more different, but I'm hoping book two will have the story turn into something of its own. Overall I enjoyed reading this story and I'm looking forward to reading book two especially after that ending.
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Reviewed in the United States on May 27, 2023
★★★★★ 5
Unraveling Fate and Fae: A Captivating Journey in "Queen of Roses"
Format: Kindle
"Queen of Roses" by Briar Boleyn is a dark fantasy romance that masterfully combines elements of myth, magic, and romance with a captivating King Arthur retelling infused with a Fae twist. From its intricately woven plot to its compelling characters, this novel delivers an immersive reading experience that will leave readers eagerly anticipating the next installment.
At its core, "Queen of Roses" is an enchanting tale of forbidden love and destiny, featuring an exceptionally slow-burn romance that ignites with the intensity of an enemies-to-lovers trope. Against a backdrop of magic and mythical creatures, the story unfolds with tension, banter, and forced proximity, drawing readers into a world filled with love, friendships, self-discovery, and betrayal.
While the novel excels in world-building, character development, and plot intricacies, some readers may yearn for a bit more fire and spice in certain aspects of the narrative. However, the promise of future developments in the series offers hope for an even more dynamic and engaging story to come. I know I personally cannot wait to get into book 2.
With a cliffhanger ending that leaves hearts racing and minds reeling, "Queen of Roses" succeeds in immersing readers from start to finish. Its dark and twisted fantasy elements are expertly balanced with moments of adventure, action, and unexpected twists, keeping readers on the edge of their seats until the very last page.
As the story delves into complex themes and explores the depths of its characters' struggles and desires, it's important to note that "Queen of Roses" may contain triggering content. Readers are advised to check the trigger warnings before diving into this captivating tale.
Overall, "Queen of Roses" is a must-read for fans of dark fantasy romance, offering a mesmerizing journey that will leave readers eagerly anticipating the next chapter in the series. With its lush prose, intricate storytelling, and unforgettable characters, this novel is sure to leave a lasting impression on all who venture into its enchanted world.
I want to extend a heartfelt shoutout to the author for granting me the opportunity to dive into "Queen of Roses" through NetGalley. It has been an absolute pleasure to explore the captivating world and characters crafted with such skill and imagination. Thank you for entrusting me with this glimpse into your enchanting world.
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Reviewed in the United States on April 1, 2024
★★★★★ 4
Right from the start, I was drawn in by the prologue!!!
Format: Paperback, Format: Paperback
Queen of Roses (Blood of a Fae #1) by Briar Boleyn
Genre General Fiction ( Adult), Romance, Sci-Fi & Fantasy, Dark Romance
“More primordial than the stars. My name was on his lips as he promised unspeakable darkness to any who came between us.”
Right from the start, I was drawn in by the prologue!!!
I’m a big fan of “touch her, and you die” vibes, but I mean, what’s also not to love about a unique Arthurian retelling with gender twists, a treacherous royal court, a dangerous quest, magical Fae & mystical monsters, entwined with a bit of spice!
Morgan, Princess of Pendrath and true heir to the throne has spent most of her life dimming her light to feel safe and to make others comfortable. She is treated as an outcast in the court and repressed by her family due to the blood of the Fae within her and forced to join the Temple of the Three as a priestess in training to one day replace Merlin.
Her brother, King Arthur, who reminds me of Joffrey from Game of Thrones, later tells her that he has other plans and offers her a choice of the Temple or to marry her off for political gain, unless… that is, she can journey through the great unknown and return with a long-lost fae weapon with enchanted powers known as Excalibur.
Her quest begins with a roguish crew that includes the mysterious, arrogant, and heart-tuggingly handsome Captain of the Royal Guard, Kairos Draven, whom she can’t decide if she wants to stab or indulge in pleasure with.
Along the way are plenty of surprises, mystical creatures, and betrayal, all while Morgan uncovers more of the truth about herself and who she can trust.
This book had intriguing storylines and lovable characters that kept me turning pages and wanting more. I can’t wait to see how it all unfolds and comes together in book 2, Court of Claws, which I just started reading!!
Read if you’re into-
Dark Fantasy/Romance
Slow–Burn
Question Everything
Magic and Action
Fae
Arthurian Legend
Stabby/Broken FFC
Morally Gray MMC
Forced Proximity
Queen of Roses is perfect for Holly Black, Jennifer L. Armentrout, and Sarah J. Maas fans.
Please check the trigger warnings page in the table of contents before reading this book.
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Reviewed in the United States on August 16, 2023