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Description
Rat TRAP ELISA KitProduct Specification Usage 1. Self prepared test equipment required for the experiment: 1, plate reader (450nm) 2, high precision sampler and gun head: 0. 5 10uL5 50uL20 200uL200 1000uL 3, 37 Incubator 4, Distilled water or deionized water 2. Sample processing and requirements: The detection range of the kit is not equivalent to the concentration range of the test substance in the sample It is recommended to estimate the concentration of the test
Product Specification
| Usage |
1. Self-prepared test equipment required for the experiment: 1, plate reader (450nm) 2, high-precision sampler and gun head: 0.5-10uL、5-50uL、20-200uL、200-1000uL 3, 37℃ Incubator 4, Distilled water or deionized water 2. Sample processing and requirements: The detection range of the kit is not equivalent to the concentration range of the test substance in the sample It is recommended to estimate the concentration of the test substance in the sample through relevant literature before the experiment and determine the actual concentration of the sample through pre-experiment. If the concentration of the test substance in the sample is too high or too low, please dilute or concentrate the sample appropriately. If the sample tested is not among the sample types listed in the instructions, it is recommended to conduct pre-experiments to verify its detection effectiveness. Serum: Whole blood samples collected in serum separation tubes were placed at room temperature 2 Hour or 2-8℃ Overnight, then 1000×g Centrifugation 20 Minutes, take the supernatant, or place the supernatant in -20℃ Or -80℃ Store, but repeated freezing and thawing should be avoided. Plasma: use EDTA Or heparin as an anticoagulant to collect a sample, and collect the sample after collection 30 Within minutes 2-8℃1000×g Centrifugation 15 Minutes, take the supernatant to detect, or place the supernatant in -20℃ Or -80℃ Store, but repeated freezing and thawing should be avoided. Tissue homogenate: With pre-cooled PBS(0.01M,pH=7.4) The tissue is flushed to remove residual blood (lysed red blood cells in the homogenate can affect the test results), and the tissue is weighed and cut into pieces. Combining the shredded tissue with the corresponding volume PBS (Generally according to 1:9 Weight to volume ratio, such as 1g The tissue samples correspond to 9mL Of PBS The specific volume can be appropriately adjusted according to the experimental needs and recorded. Recommended in PBS Add protease inhibitor) into a glass homogenizer and grind thoroughly on ice or grind in a homogenizer. For further lysis of tissue cells, the homogenate can be sonicated, or freeze-thawed repeatedly. Finally, the homogenate was mixed in 5000×g Centrifugation 5~10 Minutes, take the supernatant for detection. Cell culture supernatant: Please 1000×g Centrifugation 20 Minutes, take the supernatant to detect, or place the supernatant in -20℃ Or -80℃ Store, but repeated freezing and thawing should be avoided. Cell lysate: Precooling for adherent cells PBS Gently washed, followed by trypsinization, 1000×g Centrifugation 5 Cells were collected after minutes. The suspended cells can be collected directly by centrifugation. The collected cells were pre-cooled with PBS Washing 3 Times, every 1×10^6 Added to cells 150-200μLPBS Resuspension (recommended at PBS Adding a protease inhibitor. If the content is very low, it can be appropriately reduced PBS Volume) and the cells were disrupted by repeated freeze-thaw or sonication. The extracts were mixed in 2-8℃, 1500×g Centrifugation 10 Minutes, take the supernatant for detection. Other biological samples: 1000×g Centrifugation 20 Minutes, take the supernatant to detect. Sample Appearance: The sample should be clear and transparent, and the suspended solids should be centrifuged to remove. Sample Preservation: After sample collection, if 1 Those tested within weeks can be stored in 4℃, if it cannot be detected in time, please pack it according to the one-time usage amount and freeze it in -20℃（1 Within months), or -80℃（6 Test within months) to avoid repeated freezing and thawing. Hemolysis of the sample will affect the final test result, so hemolyzed samples are not suitable for this test. 3. Sample dilution plan: Please estimate the concentration range of the sample in advance. If your test sample needs to be diluted, refer to the dilution plan as follows: Dilution 100 Times: One step dilution. Take 5μL Sample to 495μL Within universal diluent, do 100 Double dilution. Dilution 1000 Times: Two-step dilution. Take 5μL Sample to 95μL Within universal diluent, do 20 Dilute, and then take 5μL20 Double dilute sample to 245μL Within universal diluent, do 50 Double dilution, total dilution 1000 Times. Dilution 100000 Times: Three-step dilution. Take 5μL Sample to 195μL Within universal diluent, do 40 Dilute, and then take 5μL40 Double dilute sample to 245μL Within universal diluent, do 50 Time dilution, and finally take 5μL 2000 Double dilute sample to 245μL Within universal diluent, do 50 Double dilution, total dilution 100000 Times. The amount of liquid taken during each dilution step is not less than 3μL, the dilution factor is not more than 100 Times. Each step of dilution should be mixed evenly to avoid foaming. 4. Preparations before testing: 1, please advance 10 Minutes remove the kit from the refrigerator and equilibrate to room temperature. 2, Standard gradient working solution preparation: add 1mL Universal diluent into lyophilized standard and let stand 15 Minutes until it is completely dissolved and then gently mix (The concentration is 250ng/mL). And then according to the following concentrations: 250ng/mL、125ng/mL、62.5ng/mL、31.25ng/mL、15.62ng/mL、7.81ng/mL、3.9ng/mL、0ng/mL The dilution was performed. Double dilution method: Take 7 branch EP Tubes, each tube is added 500μL Universal diluent,250ng/mL Pipette from the standard working solution 500μL To the first EP Mix evenly in a tube 125ng/mL. According to this step, suck and mix the standard working solution in turn. The last tube is directly used as a blank hole, so there is no need to suck liquid from the penultimate tube, as shown in the figure below. 3, Preparation of biotinylated antibody detection working solution: before use 15 Minutes will 100× Concentrated biotinylation detection 1000×g Centrifugation 1 Minutes, with a universal diluent 100× Concentrated biotinylated detection antibody diluted into 1× Working concentration (Example: 10μL concentrate +990μL Universal diluent), now available for use. 4, Preparation of enzyme conjugate working solution: before use 15 Minutes will 100× Concentrated enzyme conjugate in 1000×g Centrifugation 1 Minutes, with a universal diluent 100× The concentrated enzyme conjugate is diluted into 1× Working concentration (Example: 10μL Concentrate +990μL Universal diluent), now available for use. 5, 1× Wash liquid preparation: Take 10mL20× Wash liquid to 190mL In distilled water (the concentrated washing liquid taken out from the refrigerator may have crystals, which is a normal phenomenon. It can be placed at room temperature and prepared after the crystals are completely dissolved). 5. Operation steps: 1, Equilibration from room temperature 10 After minutes, remove the required slats from the aluminum foil bag, and seal the remaining slats with ziplock bag and put them back 4℃. 2, Adding samples: respectively add samples or different concentration standards according to 100μL Each well is added to the corresponding well, and the blank well is added 100μL Universal diluent. After covering the sealing film 37℃ Incubation 60 Minutes. (Recommendation: minimum dilution of sample to be tested with universal diluent 1 Times later, add the enzyme labeled plate for testing. So as to reduce the influence of matrix effect on the test results, and finally, the sample concentration needs to be multiplied by the corresponding dilution factor when calculating. It is recommended to set up double wells for all samples and standards to be tested during testing). 3, Add biotinylated antibody detection: take out the enzyme labeled plate, discard the liquid, and do not wash it. Add directly to each well 100μL Biotinylated antibiotic detection working solution, after covering with sealing film 37℃ Incubation 60 Minutes. 4, Plate washing: discard the liquid and add to each well 300μL1x Wash liquid, stand 1 Minutes, throw off the washing liquid, pat dry on absorbent paper, and repeat washing the plate 3 Times (the plate can also be washed with a plate washing machine). 5, Add enzyme conjugate working solution: add each well 100μL Enzyme conjugate working solution, after covering with sealing membrane 37℃ Incubation 30 Minutes. 6, Plate washing: discard the liquid according to the steps 4 Washing method, wash plate 5 Times. 7, Add substrate: add per well 90μL Substrate (TMB) Covered with a sealing film, 37℃ Incubation protected from light 15 Minutes. 8, Add stop solution: take out the enzyme label plate and add it directly to each well 50μL Stop solution, immediately in 450nm Wavelength measurement of each well OD Value. VI. Calculation of experimental results: Result judgment: 1, calculate the average of the standard and sample replica well OD Value and subtract the blank hole's OD Values as correction values. Taking concentration as the abscissa, OD Value is ordinate, draw the standard curve of the four-parameter logic function on the double logarithmic coordinate paper. 2, If the sample OD If the value is higher than the upper limit of the standard curve, the test should be retested after appropriate dilution and multiplied by the corresponding dilution factor when calculating the sample concentration. 7. Kit performance: 1, Repeatability: the coefficient of variation in the plate is less than 10%, the interplate coefficient of variation is less than 10%. 2, Recovery rate: added to selected healthy serum, plasma and cell culture supernatant 3 Rats at different concentration levels TRAP, calculate the recovery. 3, linear dilution: respectively in the selected 4 Healthy serum, plasma and cell culture supernatant were added with high concentration of rats TRAP, linearity was assessed by dilution within the standard curve kinetic range. |
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| Sensitivity | 1.7ng/mL | |||||||||||||||||||||||||||||||||
| Species Reactivity | Rat | |||||||||||||||||||||||||||||||||
| Theory | This kit uses double antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, HRP enzyme conjugate were sequentially added to microwells pre-coated with rat anti-tartrate acid phosphatase (TRAP) capture antibody, incubated and washed in the middle, and colored with substrate TMB. TMB was converted to blue under the catalysis of peroxidase (HRP) and to final yellow under the action of acid. There was a positive correlation between the depth of color and rat tartrate-resistant acid phosphatase (TRAP) in the sample. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450nm, and the sample concentration was calculated. | |||||||||||||||||||||||||||||||||
| Synonym | Rat Acid Phosphatase 5, Tartrate Resistant (TRAP) ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Tartrate-resistant acid phosphatase (TRAP), also called ACP5, is a glycosylated monomeric metalloproteinase expressed in mammals. It has a molecular weight of about 35kDa, has an alkaline isoelectric point (7.6-9.5) and is optimal for activity under acidic conditions. TRAP is synthesized as a latent proenzyme and activated by proteolysis and reduction. It is distinguished from other mammalian acid phosphatases by its resistance to tartrate inhibition and its molecular weight. | |||||||||||||||||||||||||||||||||
| Storage Temp. | Unopened kit, stored at 4 °C, shelf life 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 3.90-250ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.5 ★★★★★
Based on 29 reviews
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Product Reviews
★★★★★ 4
Slow, sorry but good
Format: Kindle
This was a slow moving book. Lots of character pov chapters, lots of superfluous descriptions and endless courtly appearances stalled this book to start. Once you get into the heart of the story, it takes off. Before you know it the book is done. My favorite character is Reyna. She is so strong. She is true to herself. She gets into a lot of trouble with her headstrong ways but it's entertaining. I have high hopes for Lorcan. He is honorable to a fault. Thane had turned out to be better than I thought but i still don't like him. Eislin is useless. Great plot twists at the end. I'm looking forward too book 2.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 6, 2020
★★★★★ 5
Very detailed! Beautiful world building! Strong Heroine!
Format: Kindle
Starts off a little slow and confusing with different POV’s, but starts to all come together towards the middle to make an elaborate plot line and makes it all worth it. Beautiful world building and attention to detail as well as great writing. The cliffhanger was gut wrenching! Can’t wait for the next book!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 4, 2020
★★★★★ 4
Interesting
Format: Kindle
This was a very captivating book once you got into it thoroughly. But the third person perspective was a bit hard to get used to. But as you got into it and followed the different characters, it was interesting and filled with intrigue, conflict and forbidden love.
I can’t wait to read the next one and to complete the series.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 12, 2022
★★★★★ 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.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 16, 2023
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