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Description
Human DNMT1 ELISA KitProduct Specification Usage 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, Sample handling and requirements: Tissue homogenate: with pre cooled PBS 0. 01M,pH=7. 4 ) Rinse the tissue to remove residual blood (lysed red blood cells in the homogenate will affect the measurement result),
Product Specification
| Usage |
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, Sample handling and requirements: Tissue homogenate: with pre-cooled PBS （ 0.01M,pH=7.4 ) Rinse the tissue to remove residual blood (lysed red blood cells in the homogenate will affect the measurement result), and break the tissue after weighing. Combine the fragmented 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 ground thoroughly on ice. 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 lysate: pre-cooling 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 150-200uL PBS 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 fluids: 1000xg Centrifugation 20 Minutes, take the supernatant to detect. 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 10ng/mL ） And then according to the following concentrations: 10ng/mL 、 5ng/mL 、 2.5ng/mL 、 1.25ng/mL 、 0.625ng/mL 、 0.3125ng/mL 、 0.15625ng/mL 、 0ng/mL The dilution was performed. Double dilution method : Take 7 branch EP Tube , added to each tube 500uL Universal diluent ,10ng/mL Pipette from the standard working solution 500uL To the first EP Mix evenly in a tube 5ng/mL Standard Working Solution , according to this step, absorb and mix evenly in turn. The last tube is directly used as a blank hole , 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 Min. The concentrated biotinylated antibody was concentrated in 1000×g Centrifugation 1 Minutes, with a universal diluent 100× The concentrated biotinylated antibody was diluted into 1× Working concentration (ex: 10uL Concentrate +990uL 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× concentrate HRP The enzyme conjugate is diluted into 1× Working concentration (ex: 10uL Concentrate +990uL Universal Diluent) , now available for use. 5 、 1× Wash liquid preparation: Take 10mL 20× Wash liquid to 190mL In distilled water (the concentrated washing liquid taken out of the refrigerator may have crystals, which is a normal phenomenon. It can be left at room temperature and prepared after the crystals are completely dissolved). 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 100ul Each well is added to the corresponding well, and the blank well is added 100uL 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: take out the enzyme plate, discard the liquid without washing. Directly add biotinylated antibody working solution to each well 100uL , after covering the sealing film 37℃ Incubation 60 Minutes. 4 Plate washing: discard the liquid and add to each well 300uL 1x 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 Adding enzyme conjugate working solution: adding enzyme conjugate working solution to each well 100uL , after covering the sealing film 37℃ Incubation 30 Minutes. 6 Plate washing: discard the liquid according to the steps 4 Washing method, wash plate 5 Times. 7 Substrate addition: substrate is added per well ( TMB ） 90uL Covered with a sealing film, 37℃ Incubation protected from light 15 Minutes. 8 Add stop solution: take out the enzyme label plate and directly add stop solution to each well 50uL , immediately in 450nm Wavelength measurement of each well OD Value. 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. |
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| Theory | This kit uses double antibody sandwich enzyme-linked immunosorbent assay (ELISA). To the microwells pre-coated with DNA Methyltransferase 1 (DNMT1) capture antibody, sample, standard product, biotin-labeled detection antibody, and HRP enzyme conjugate were added in sequence, incubated and washed in the middle, and colored with substrate TMB. TMB is 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 DNA Methyltransferase 1 (DNMT1) in the sample. The absorbance (0D value) was measured with a microplate reader at a wavelength of 450nm, and the sample concentration was calculated. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human DNA Methyltransferase 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | DNA methyltransferase 1 (DNMT1) is an enzyme that catalyzes the transfer of methyl groups to specific CpG structures in DNA, a process called DNA methylation. It is encoded by the DNMT1 gene. It is part of the DNA methyltransferase family, which consists primarily of DNMT1, DNMT3A, and DNMT3B. The enzyme is responsible for maintaining DNA methylation, thereby ensuring the fidelity of this epigenetic pattern in cell division. However, it can catalyze novel DNA methylation in specific genomic settings, including transposable elements and paternal imprinted control regions. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Carry out incubation in strict accordance with the specified time and temperature to ensure accurate results. All reagents must reach room temperature 20-25 °C prior to use. Store reagents in refrigeration immediately after use. 2. Incorrect plate washing may lead to inaccurate results. Make sure to drain the liquid from the wells as much as possible before adding the substrate. Do not allow the wells to dry out during incubation. 3. Eliminate the residual liquid and fingerprints at the bottom of the plate, otherwise it will affect the OD value. 4. The substrate color development solution should be colorless or very light in color, and the substrate solution that has turned blue cannot be used. 5. Avoid cross-contamination of reagents and specimens to avoid wrong results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Any reaction reagent cannot come into contact with the bleaching solvent or the strong gas emitted by the bleaching solvent. Any bleaching component will destroy the biological activity of the reaction reagents in the kit. 8. Expired products cannot be used, and components with different item numbers and batch numbers cannot be mixed. 9. Recombinant proteins from sources other than the kit may not match the antibodies in this kit and are not recognized. 10. If the disease may be spread, all samples should be managed well, and the samples and testing devices should be handled according to the prescribed procedures. |
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| Storage Temp. | Store the unopened kit at 4 °C. Shelf life: 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.156-10ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates and other biological fluids |
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4.8 ★★★★★
Based on 15 reviews
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Product Reviews
★★★★★ 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.
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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.
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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