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Description
Human C5aR 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 specimens using EDTA or heparin as anticoagulants and centrifuge them at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. The supernatant can be tested or stored at -20℃ or -80℃, but repeated freezing and thawing should be avoided. 3. 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 Complement component 5a receptor (C5aR). 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 Complement component 5a receptor (C5aR) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Complement component 5a receptor ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Complement component 5a receptor 1 (C5aR1), also known as CD88 (cluster of differentiation 88), is a G protein-coupled C5a receptor. It functions as a complement receptor. The C5a receptor regulates inflammatory responses, obesity, development, and cancer. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, and other biological fluids |
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4.0 ★★★★★
Based on 1416 reviews
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Product Reviews
★★★★★ 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
★★★★★ 3
3.5 stars, A little boring to say the least.
Format: Kindle
Wow so I'm not sure where to begin on this one. This was a very different take on the legend of Arthur and Excalibur. This is told from the point of view of Morgan the sister of Arthur. Honestly the first 50% of this book is world building and character building which unfortunately was super boring for me. Morgan to me was a female MC that had a hard time in believing in herself. Sometimes taking too long to understand exactly what was going on around her. Draven was also a different male MC, like I couldn't put my finger on him and what he was all about. It was not until the last 10% of the book did we get some answers on the mystery that is Draven. The other 50% of the book centered around this big journey with everyone having a different motive. We see a spark of magic around this time that had me excited but then we never expanded upon that and what it could mean for the female MC. I feel like I want to read the second book just to see where this goes, but the spice was probably a 2 out of 5. Side characters are ok, Lancelet was fun but I almost felt like I wanted more.
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Reviewed in the United States on September 13, 2023
★★★★★ 5
A bewitching retelling of Arthurian legend!
Format: Kindle
In a land where the Fae have nearly become only a legend and those who still posses even a morsel of the blood are few and far between, Morgan finds herself cast aside by most of society due to her rumored half-Fae lineage, including her brother, King Arthur. With the kingdom at the brink of war, Arthur entrusts her with a quest to retrieve a Fae weapon of legendary power: the sword of Perun, Excalibur. Accompanied by men she loathes, Captain Kairos Draven and Ragnar Whitehorn, she embarks on her long and unbeknownst perilous journey, only to find that things she once believed to be myth are in fact very real. With devastating twists, omitted truths, witty banter and fierce action, Queen of Roses leaves you begging to know more about the secrets of Aercanum!
Wow, wow, wow! Going into this story, I did not realize that it was going to be a retelling of Arthurian legend, especially not one with a fantastical twist! The unique spin almost gave me The Witcher vibes and I think adding Fae into the mix was quite interesting. I knew the basics of the legend but after reading this book, it has piqued my interest and makes me want to learn more about it. My attention was snatched as soon as I finished the prologue and I knew that I was going to devour this story. I truly enjoyed the gender swaps and even how Arthur was portrayed as villainous. Morgan’s past and even her parts of her present is absolutely heartbreaking, and I felt for her at times. I can only recall one other book that made me hate characters the way I despised Florian and Arthur, leaving me with my blood boiling and feeling disgusted. Even after finishing the book, Draven is still a mystery to me and I cannot figure out how to feel about him. I guess they just means that the author did an excellent job at conveying each character’s persona! The rich world building and imagery made it easy for me to visualize the places that the group visited along their journey. I am truly engulfed in this story and I cannot wait to see wait fate awaits Morgan and how the Fae will be even more incorporated in the next book!.
I received a free copy of this book and am voluntarily leaving a review.
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Reviewed in the United States on August 11, 2023
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