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Description
Rat 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: 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 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 | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.6 ★★★★★
Based on 18 reviews
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Product Reviews
★★★★★ 4
Great debut!
Format: Hardcover
“I will decimate anyone who tries to hurt you; anyone who lays a hand on you.”
DV awareness
History erasure
Magic suppression
Found family
Chosen one
Slow burn to spice
I was happy that there’s a heavy exposure to domestic violence awareness and that Cin found the will and courage to acknowledge and get out of the abusive relationship she was in. She is really working toward taking back her voice, her choice and her power. The story covers themes of loss, betrayal and oppression as well as the power of friendship and having people truly see you. And Landers, our mmc, delicious.
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Reviewed in the United States on June 18, 2025
★★★★★ 5
Emotional & Powerful Read!
Format: Paperback, Format: Paperback
The Book of Cin by HM Wolfe was such an emotional and powerful read! As the first installment in The Stories Trilogy, it completely hooks you and leaves you desperate for more. I absolutely loved watching the FMC slowly grow into her abilities after years of abuse and struggling with her confidence. It felt raw and real in a way that stuck with me 🖤
This story has everything.. found family, betrayal, and moments of truly heart-wrenching grief. But what I loved the most was the belief that love and chosen family can help heal even the deepest, unseen wounds. Seeing that healing unfold made this journey incredibly rewarding ✨
If you love fantasy with emotional depth, an amazing magic system, character growth, and meaningful relationships, you NEED to pick this one up! 📚🔥
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 23, 2026
★★★★★ 4
Well…
Format: Kindle
I enjoyed the book, it had an interesting plot and magic system. The book is full of smirks, hissing, and crying. OMG the crying! The FMC cries from front to back of the book! I absolutely loved Daggermouth and read this book bc that, but I feel like she had a different editor maybe?! Idk what happened, but this book would have been so much better without the MMC smirking on every page that the FMC was crying on.
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Reviewed in the United States on May 29, 2026
★★★★★ 5
Top Book of 2024
Format: Kindle
May Contain Spoilers!
Where do I even begin?
I'm still in shock of how amazing this book was, how it touched my heart, and now how I have to wait for H. M. Wolfe to write the second. Because those last few chapters...went from 0 to 100 quick. So much new information, new relationships and loss, that now the tides have turned, and we NEED more.
This is a fantasy book of love, loss, the power of friendship, and finding one owns strength. TBOC is about Hyacinth whose home is at Asrai’s Academy for Orphans. She was torn between the safety of home or leaving it all behind for a chance at a new life. But when a mysterious messenger appears, whispering rumors of a God among Fae, everything Hyacinth thought she knew about the Realms comes crashing down, and she must now make a new journey.
This book ate me up and spit me back out. I was not anticipating the emotions I would feel within these pages. I'm talking I was laughing, then I was swooning, then I was on the edge of my seat, then I'm ready to slap someone, then I'm rooting so hard for certain characters I don't wan to stop, then my heart is broken, then I'm excited af.... it was all an emotional rollercoaster. The last few chapters contained a lot of new information, so of course as much as it doesn't leave off on a complete cliffhanger, I was over here like scraping the bottom of an ice cream cup, wanting more.
The magic system starts out ambiguous, but as you continue reading you start to get more and more to build off of and start to piece together the world H. M. Wolfe created. TBOC brings in eight main characters that test alliances, grow in friendships, fall in love, become a team, and even break bonds.
The friendship among the girls was everything. Ata is a spit fire, Pria is sweet yet terrifying. The banter and protectiveness these three had for one another made me want to join in with them for breakfast. I liked how there was multiple romances going on at once. Yes we have the main characters, but it just wasn't always about them, and it was great to see how others shifted and grew too.
And last the romance, I've never rooted so hard for two characters. We're talking slow burn between them. In the beginning you're not sure how this is all going to play out, but Landers is a tatted, "touch her and die" kind of MMC, which who cannot love that? Hyacinth definitely had to grow within the chapters and learn how to be strong, because sometimes she was making me mad and I just wanted to step in there for her. But Landers was everything she needed and it was so well written, which is why by time you get to the end you only want more of them!
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Reviewed in the United States on September 24, 2024
★★★★★ 3
cliffhanger, no 2nd book in sight
Format: Kindle
The book itself was pretty good. The plot is unique and full of suspense and action, although some of it felt redundant. I particularly enjoyed the realistic growth cycle of the female lead from abuse survivor with low self-esteem to badass warrior.
I never would have read it though if I had known this is not complete, and there is no information available on the next book.
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Reviewed in the United States on November 17, 2024