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Description
Rat ERK1 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. 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 40 ng/mL). Then dilute to the following concentrations: 40 ng/mL, 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 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 40 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 20 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 utilizes 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 an extracellular signal regulated kinase 1 (ERK1) capture antibody. After incubation and washing, the assay 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 extracellular signal regulated kinase 1 (ERK1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Extracellular Signal Regulated Kinase 1ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Extracellular signal-regulated kinase 1 (ERK1), also known as p44MAP, is an enzyme encoded by the MAPK3 gene. The protein encoded by this gene is a member of the mitogen-activated protein kinase (MAP kinase) family. MAP kinases, also known as extracellular signal-regulated kinases (ERKs), function in a signaling cascade that regulates various cellular processes, such as proliferation, differentiation, and cell cycle progression, in response to various extracellular signals. Activation of this kinase by upstream kinases leads to its translocation to the nucleus, where it phosphorylates its nuclear targets. Alternatively spliced transcript variants encoding different protein isoforms have been described. | |||||||||||||||||||||||||||||||||
| 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 | 0.625-40 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.2 ★★★★★
Based on 1434 reviews
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Product Reviews
★★★★★ 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
★★★★★ 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
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