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Description
Human PKB ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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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| Sensitivity | 0.14 ng/mL | |||||||||||||||||||||||||||||||||
| 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 protein kinase B (PKB) capture antibody. After incubation and washing, the assay 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 protein kinase B (PKB) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Protein kinase B ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Protein kinase B (PKB), also known as Akt, is a collective term for a group of three serine/threonine-specific protein kinases that play key roles in multiple cellular processes, such as glucose metabolism, apoptosis, cell proliferation, transcription, and cell migration. Three distinct genes encode protein kinase B isoforms, designated AKT1, AKT2, and AKT3, encoding RAC α, β, and γ serine/threonine protein kinases, respectively. The terms PKB and Akt may refer collectively to all three genes but are sometimes used to refer to PKB alpha and Akt1 individually. Akt isoforms are overexpressed in various human tumors and are genomically amplified in gastric adenocarcinoma (Akt1), ovarian cancer (Akt2), pancreatic cancer (Akt2), and breast cancer (Akt2). | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.1 ★★★★★
Based on 5 reviews
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Product Reviews
★★★★★ 4
Interesting story
Format: Paperback
The Alchemist is an engaging modern fable centered on self-discovery, personal destiny, and following your dreams. It offers interesting life lessons and an easy-to-read story. While its themes draw on various spiritual ideas and may feel unconventional, it still delivers meaningful reflections on purpose and perseverance. Overall, a thought-provoking and enjoyable read.
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Reviewed in the United States on April 26, 2026
★★★★★ 5
One Of The Most Important Books Ever Written
Format: Paperback
The Alchemist by Paulo Coelho is in my opinion, one of the most important books ever written. It's no wonder that it's one of the best-selling books of all time with more than 150 million copies sold. Those numbers are in spite of and defiance to the author's encouragement to file share his now legendary novel.
But sales statistics are not the criterion on which I stake this claim. It is the impact on the lives of those that have read it, and the impact on the lives that have not yet been born, that I boldly state the above.
The simplicity of its unfoldment may at first belie the depth of its wisdom. But the simplicity is part of the profundity that allows the reader to assimilate the messages without resistance.
The first time I read The Alchemist;, I read it cover to cover in one sitting. No bathroom break, no cup of water pause, no telephone distractions. I read it from the first word to the last in what seemed like a single breath.
I simply couldn't put it down.
I was swept, swung, and swallowed up by this extraordinary narrative. The result was transformation. I was not the same man that began reading the novel by the time I finished it.
I would thereafter read it again and again. The messages are so layered that with every new pass comes new understanding and enlightenment.
At the core of the novel is the fulfillment of one’s destiny. There is a force in the Soul of the World that wants us all to live out our personal legend. When we truly desire something, the entire Universe conspires to help us achieve it. With love, all things are possible. With love, we can perform miracles.
These themes are artfully integrated and clothed within a modern day fable. They come in varying iterations from a string of colorful characters that propel the protagonist towards a profound and satisfying ending.
The Alchemist is the story of Santiago, a shepherd boy from Spain that crosses the desert of Africa in search of a hidden treasure. It was revealed to him in a reoccurring dream that the treasure is buried by the Ancient Pyramids in Egypt.
What he naively believes will take three weeks to complete becomes a journey that expands nearly three years. After miles of traveling through the desert, the boy arrives at an oasis where he falls in love with a beautiful desert woman named Fatima. The boy is willing to abandon his dream of finding his treasure for the procurement of her love.
But one night, beneath the light of a full moon, he meets a man straddled upon a mighty horse. This elusive man is known in legend and folklore as The Alchemist.
The Alchemist is a man imbued with profound wisdom and mystical powers. He is reportedly over 300-years-old and can turn lead into gold.
He reminds the boy that his destiny is to find the treasure. That he must continue. If it is true love, Fatima will be waiting for him after his destiny is fulfilled.
Reluctantly, the boy continues his journey deeper into the desert accompanied by The Alchemist. But the desert is at war, and they are soon captured by Bedouin sentinels and accused of spying for the enemy.
For this crime, they are sentenced to death.
The Alchemist saves them temporarily by proclaiming that the boy is an Alchemist. He tells the general that if he wanted to, the boy could turn himself into the wind and destroy their entire camp. The general is intrigued! He wants to see this happen. He promises to spare their lives if it is done. He grants them three days.
But how does a shepherd boy from Spain turn himself into the wind?
Paulo Coelho says that he wrote “The Alchemist” in two weeks. It is almost hard to imagine that something so far-reaching could have been conjured up in two weeks time. He said it was his own journey that he was writing down, parabolically speaking.
The impact of “The Alchemist” comes from the realization that Santiago’s journey is Everyman’s journey. His story is our story. If we tap into the Soul of the World and dare to live out our destiny, we will discover that within us all is The Alchemist.
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Reviewed in the United States on April 21, 2017
★★★★★ 5
A must read
Format: Kindle
A must read for everyone even remotely interested in self fulfilment and awakening. Can be read in an afternoon and echo for a life time.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 25, 2026
★★★★★ 5
A Beautiful Edition of a Timeless Journey
This deluxe edition transforms Coelho's beloved fable into a truly special keepsake. The larger format, quality paper, and thoughtful design details make it feel like the gift it's meant to be—whether you're giving it to someone at a crossroads in their life or treating yourself to an upgrade from a worn paperback copy.
The story itself remains as powerful as ever. Santiago's journey across the desert in search of treasure is deceptively simple on the surface, but Coelho's allegory works on multiple levels. Yes, it's about following your dreams, but it's also about learning to read the signs the universe offers, understanding that the journey itself transforms us, and recognizing that what we seek is often closer than we think.
I know some readers find the philosophy too simplistic or New Age-y, but I think that misses the point. Coelho isn't trying to be subtle—he's distilling universal truths into their essence. Sometimes we need straightforward wisdom delivered clearly, especially when we're overthinking our own paths.
This edition makes a perfect graduation gift, retirement present, or milestone marker for anyone embarking on something new. The story has earned its place as a modern classic, and this deluxe version does it justice.
"When you want something, all the universe conspires in helping you to achieve it." Still resonates.
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Reviewed in the United States on January 20, 2026
★★★★★ 5
Reading this book was an enlightening experience for me.
Format: Kindle
The Alchemist by Paulo Coelho provided hours of magical reading. It isn’t a plot-driven book, but rather a novel that provides a quiet sense of purpose and a deeper understanding of the universe. The period the story takes place isn’t clear, but it begins in Spain.
Santiago, a young man, perhaps in his teens, studied to be a priest, but realized what he really wanted to do was to be a shepherd, to move around, see new places. With the blessings of his family, he sets out with his flock of sheep and discovers not only the world, but himself. It soon becomes clear that what he searches for is an alchemist, a person who transforms things for the better.
Santiago meets people along the way who teach him many truths, some good, some evil, but each experience leads him closer to life’s true meaning.
This novel has an interesting history. “The Foreword” written by the author in 2014 speaks about the interesting path the novel has taken. The book was first published in 1989 in his native Brazil. Only one person bought a copy the first week of its release, then six months later another copy sold to the same person! His publisher cancelled the contract. Another publisher agreed to take the book and gradually had great success. An American read the book and wanted to translate the book from Spanish to English and find a publisher in the United States. It didn’t happen overnight, but the book became a phenomenon, has been translated into more than eighty different languages, and has sold thousands of copies.
Paulo Coelho never wavered in his faith in the book because it speaks of his own beliefs: A man sets out on a journey seeking a beautiful or magical place and then realizes the treasure lies within himself.
Reading The Alchemist was an enlightening experience for me. I highly recommend it for those who appreciate life’s purpose and who seek wisdom of the universe.
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Reviewed in the United States on October 7, 2025