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Description
Human Rac1 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: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled PBS and resuspend 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. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 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 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 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 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 Ras-Related C3 Botulinum Toxin Substrate 1 (Rac1) capture antibody. After incubation and washing, the sample 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 Ras-Related C3 Botulinum Toxin Substrate 1 (Rac1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Ras Related C3 Botulinum Toxin Substrate 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Ras-related C3 botulinum toxin substrate 1, also known as Rac1, is a protein encoded by the RAC1 gene. This gene can produce multiple alternatively spliced forms of the Rac1 protein, which appear to have distinct functions. It is a small signaling G protein (more specifically, a GTPase) and a member of the Rac subfamily of the Rho family of GTPases. Members of this superfamily appear to regulate a variety of cellular events, including control of GLUT4 translocation to glucose uptake, cell growth, cytoskeletal reorganization, antimicrobial cytotoxicity, and protein kinase activation. It is a pleiotropic regulator of many cellular processes, including the cell cycle, cell adhesion, motility, and epithelial differentiation. It is ubiquitously expressed in tissues and drives cell motility by forming lamellae. It is abundantly expressed in insulin-sensitive tissues, such as adipose tissue and skeletal muscle. It is also required for glucose uptake in skeletal muscle, which is activated by exercise. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids |
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★★★★★ 5
A Great Graduation Gift
Format: Hardcover
This was one of my favorite books when I was a child. I've been purchasing this as a graduation gift for years. I gave it to both my daughters when they graduated high school and was happy to read that very copy we gifted our daughter to my granddaughter. The book has also helped me in some difficult times of my own life. Dr. Seuss is timeless and this makes a great gift.
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Reviewed in the United States on May 12, 2026
★★★★★ 5
The Perfect Gift for a New Beginning
Format: Hardcover
Oh, the Places You'll Go! by Dr. Seuss is a timeless and encouraging read filled with humor, wisdom, and inspiration about life’s journey and all of its ups and downs. This is such a meaningful gift for a high school senior, graduate, or anyone stepping into a new chapter of life.
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Reviewed in the United States on May 7, 2026
★★★★★ 5
I recommend using this book. Read review.
Format: Hardcover
I recommend this book. Not only because it's a Dr. Sue's book, but because at the end of the school year. I ask my son's teacher to sign it. It's a sweet memory to have and it will be a sweet memory to look at once they graduate Elementary. It's a must. The price is great. Quality... Everything about it is great. Most of all.... Reading the their teacher's have to say about them. Is so sweet!
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Reviewed in the United States on May 23, 2026
★★★★★ 5
Dr. Suess. Enough Said.
Format: Hardcover, Format: Hardcover
Welcome to the whimsical world of Dr. Seuss, where "Oh, the Places You'll Go!" isn't just a book—it's a roadmap to life's most thrilling escapades! If life were a rollercoaster, this book would be the front-row seat to the wildest ride of all.
From the moment you crack open its vibrant pages, you're whisked away on an exhilarating journey filled with colorful landscapes, quirky characters, and nuggets of wisdom disguised in Seussian rhymes. It's a literary wonderland where imagination reigns supreme, and the words dance off the pages, pulling you into an adventure unlike any other.
Dr. Seuss, the maestro of whimsy and wonder, takes you on a whirlwind exploration of life's ups and downs, encouraging you to embrace every twist, turn, and unexpected detour with unbridled enthusiasm. Whether you're just starting out on life's path or navigating its winding roads, this book serves as a beacon of hope, inspiration, and boundless possibilities.
Now, let's talk about the humor woven into every verse—oh, it's a riot! Dr. Seuss's playful language and zany illustrations transform life's perplexities into delightful puzzles to solve. You'll find yourself chuckling at the absurdity of the situations and nodding knowingly at the profound truths hidden within the jest.
And oh, the illustrations! They're a visual feast for the eyes, bursting with vibrant hues and fantastical landscapes that transport you to worlds beyond your wildest dreams. The whimsical characters and their expressions evoke emotions that range from sheer joy to contemplative reflection, leaving an indelible mark on your heart.
But wait, there's more! This book isn't just for kids—it's a timeless gem that resonates with readers of all ages. Whether you're 8 or 80, the wisdom embedded in its verses speaks volumes about the human spirit's resilience, determination, and unwavering courage to conquer life's myriad challenges.
In essence, "Oh, the Places You'll Go!" isn't merely a book; it's a celebration of life's rollercoaster, a symphony of laughter, and a profound reminder that the journey itself is the greatest adventure of all. So, buckle up, embrace the whimsy, and get ready to soar to unimaginable heights—you're in for an exhilarating ride!
*Disclaimer: May induce spontaneous outbreaks of giggles and an insatiable thirst for adventure!*
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Reviewed in the United States on November 15, 2023
★★★★★ 5
Oh, the places you'll go! Book Review
Format: Hardcover
Perfect hardcover book for such an affordable price!
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Reviewed in the United States on May 25, 2026