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Description
RNA Binding Protein Immunoprecipitation (RIP)KitProduct Specification Usage 1, sample cracking 1. 1 animal cell (1) to prepare 405 ul Complete RIP Lysis Buffer (400 ul RIP Lysis Buffer + 4 ul Protease Inhibitor (100 x) + 1 ul RNase Inhibitor); (2) 2xl07 cell samples were collected and washed by adding 2 mL PBS. After centrifugation, the supernatant was discarded to collect cell precipitation, and 1 4 cell samples were reserved for RNA extraction and purification in Input group (stored in nuclease
Product Specification
| Usage | 1, sample cracking 1.1 animal cell (1) to prepare 405 ul Complete RIP Lysis Buffer (400 ul RIP Lysis Buffer + 4 ul Protease Inhibitor (100 x) + 1 ul RNase Inhibitor); (2) 2xl07 cell samples were collected and washed by adding 2 mL PBS. After centrifugation, the supernatant was discarded to collect cell precipitation, and 1/4 cell samples were reserved for RNA extraction and purification in Input group (stored in nuclease-free EP tube at -80°C). (3) 400uL Complete RIP Lysis Buffer was added to the remaining cell precipitate and resuspended, and the precipitate was blown and mixed 10 times with a pipettes gun to fully lyse the cells. The cells were bathed in ice for 30min, vortexed every 5min, 10s/ time. 20% power, ultrasonic 3 s, intermittent 3 s. After centrifugation at 12000rpm for 10min at 4 °C, the supernatant was removed, recorded as Lysis, and divided into triplicate according to 150uL (IP), 150uL (lgG), and 100uL (Input), and the Input sample was stored at -80°C until use. 1.2 animal tissues (1) to prepare 405 ul Complete RIP Lysis Buffer (400 ul RIP Lysis Buffer + 4 ul Protease Inhibitor (100 x) + 1 ul RNase Inhibitor); (2) grinding: take fresh organizations or low temperature (about 0.3 g), put in precooling of mortar after sterilization, to join the liquid nitrogen quick grinding to powder, and collect about a quarter of the tissue samples reserved for subsequent Input set of RNA extraction and purification (using EP pipe storage without nuclease, - 80 ° C). (3) to join in mortar samples 250 ul Complete RIP Lysis Buffer, continue to grind on the ice samples into 5-10 min to the exquisite homogenate, transferred to the new EP in the tube, Then add the remaining 150uL Complete RIP Lysis Buffer to the mortar to collect the remaining samples, and transfer them to the EP tube as well. (4) The EP tube containing the sample homogenate was fully lysed for 30min on ice, vortexed every 5min, 10s/ time. After the lysis was completed, the cell ultrasonic breaker was used for 8-10min in an ice bath, with 20% power, ultrasonic 3s, intermittent 3s. (5) Centrifugation at 12000rpm for 10 min at 4°C, the supernatant was removed, and then the volume of RIP Lysis Buffer was added to the supernatant to 400uL, mixed, and divided into three parts according to 150uL (IP), 150uL (lgG), and 100u L (lnput). Input samples were stored at -80°C until use. 2, preparation of magnetic beads (1) Two 1.5mL EP tubes were prepared and labeled with lgG tube and IP tube, respectively. The original ProteinA/G Magnetic Beads tube was upside down 10 times. After the magnetic beads and liquid were mixed, 30uL were taken out into lgG tube and IP tube, respectively. (2) for every 300 ul tube join RIP for Wash Buffer, pipetting gun beat five blending, placed in a magnetic rack let stand for 1 min, abandon in qing dynasty, the three steps; (3) The beads were resuspended in 300 ul of RIP Wash Buffer. 3, magnetic beads in combination with antibodies (1) 3-5ug target antibody was added into IP tube, and 3-5ug target antibody of the same host lgG was added into lgG tube, and then incubated in a silent mixer for 2 hours at room temperature. (2) Put the two tubes on the magnetic rack for 1min, and discard the supernatant; (3) to join 300 ul RIP for Wash Buffer, 5 times, percussion and mixed with pipetting gun in magnetic rack let stand for 1 min, in qing dynasty, the steps for 3 times. 4, RNA binding protein immunoprecipitation (1) Prepare 1.8mL RIP Buffer (1720uL RIP Wash Buffer+70uL 0.5M EDTA+10uL RNase Inhibitor); (2) the lgG obtained from step 3 respectively added into the pipe and the IP 900 ul RIP Buffer, 150 ul Lysis obtained from step 1 on mute mixer, 4 ℃ incubation for the night; (3) put the two tubes in magnetic rack let stand for 1 min, abandon the supernatant; (4) Add 300uL RIP Wash Buffer to each tube, blow and mix 5 times with a pipetting gun, stand on a magnetic rack for 1 min, discard the supernatant, and this step is performed 5 times. (5) will join the 300 ul RIP each tube for Wash Buffer, pipetting gun beat five blending, take 100 ul mixture to the new tube, marked as (1) tube, the remaining liquid for (2) pipe, the 4 tubes placed in a magnetic rack let stand for 1 min, abandon supernatant, (1) tube plus 100 ul Elution Buffer, After boiling for 10min, the supernatant was taken to a new tube, and 10uL of 6×Loading Buffer was added to mix for WB detection. ② The tube was used to purify RNA, and the grouping Settings are shown in the table below:
(1) step 1 reserved Input samples and lgG - (2) pipe, IP - (2) each to join 500 ul Trizol, vortex blender, let stand at room temperature for 5 min, then add 100 ul chloroform, vortex blending, 4 ° C 14000 RPM centrifuge for 10 min, taking the upper water phase (about 300 ul) to the new tube, marking; (2) Add 50uL Salt Solution, mix with 550uL isopropanol, let stand at -80°C for 2-4h (or let stand overnight), place at 4°C to thaw before use; (3) 4 ° C 14000 RPM centrifuge for 10 min, abandon the supernatant; (4) 500uL of 75% ethanol was added, centrifuged at 4°C at 14000rpm for 10min, and the supernatant was discarded. This step was repeated 3 times. (5) Open the cover and dry at room temperature, add 10-20uL DEPC H2O, blow with pipetting gun to completely dissolve the RNA, and store at -80°C for later use. |
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| Synonym | RNA Binding Protein Immunoprecipitation (RIP)Kit | ||||||||||||||||||||||||||||||||||||
| Description |
1. Experimental principle RIP (RNA Binding Protein lmmunoprecipitation) is a technique for studying the binding of RNA to proteins in cells. It is a powerful tool to understand the dynamic process of post-transcriptional regulatory network. This technology mainly uses the specific antibody of the target protein to precipitate the corresponding RNA-protein complex, and then after isolation and purification, the RNA bound to the complex is verified by q-PCR or high-throughput sequencing.
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| Composition |
Note: 1, need to bring your own Trizol, 75% ethanol, chloroform, isopropyl alcohol, reverse transcription kit and fluorescent dye; 2, 6 t for a single set (1 set of IP or 1 set of IgG) immune precipitation experiment, the operation steps behind the 1 set contains the IgG and IP, it takes 2 t reagent. |
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| General Notes | The kit is mainly used in animals. | ||||||||||||||||||||||||||||||||||||
| Storage Temp. | Each component was stored according to the corresponding storage temperature, and the validity period was 1 year. |
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4.8 ★★★★★
Based on 18 reviews
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Product Reviews
★★★★★ 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
★★★★★ 4
Enchanting
Format: Kindle
"Queen of Roses" by Briar Boleyn is a delightful and refreshing reimagining of the classic tale of King Arthur, with a captivating twist that places the spotlight on Morgan, a character who has often been overshadowed in traditional retellings. Boleyn's creative decision to shift the narrative perspective to Morgan breathes new life into the story, offering readers an intriguing and compelling look at the Arthurian world from an entirely different angle.
One of the most commendable aspects of this book is its incorporation of Fae elements, which adds an enchanting layer of magic and mystery to the already familiar Arthurian setting. Boleyn skillfully weaves the world of the Fae into the narrative, creating a captivating backdrop against which the events of the story unfold. This addition not only adds depth to the world-building but also provides ample opportunities for twists and turns that keep readers thoroughly engrossed.
However, while the book boasts numerous strengths, it does have one noticeable flaw: the characterization of Morgan. While it is reasonable to create a flawed and complex protagonist, it appears that at times, Morgan's character becomes overly difficult and hard to relate to. Her persistently negative perception of one of the main male characters, who is a potential love interest, despite his efforts to support and assist her, may come across as somewhat irrational and could test the patience of some readers. Striking a balance between a strong, independent character and one who can recognize genuine support and affection could have enhanced the overall reader experience.
Nonetheless, the allure of "Queen of Roses" lies in its innovative approach to the Arthurian legend and its skillful blending of fantasy elements into a familiar narrative. Boleyn's evocative prose draws readers into a world where magic, destiny, and fate entwine, leaving us eager to uncover the mysteries that unfold within the pages.
I received a free copy of this book via Booksprout and am voluntarily leaving a review.
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Reviewed in the United States on July 28, 2023
★★★★★ 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
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