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Description
Rat Cyclin-E1 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 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 a G1/S-specific cyclin-E1 (Cyclin-E1) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of G1/S-specific cyclin-E1 (Cyclin-E1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat G1/S-specific cyclin-E1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | G1/S-specific cyclin E1 is a protein encoded by the CCNE1 gene. This gene encodes a member of the highly conserved cyclin family, whose members are characterized by dramatic cyclical protein abundance during the cell cycle. Cyclins function as regulators of CDKs. Different cyclins exhibit distinct expression and degradation patterns, contributing to the temporal coordination of each mitotic event. This cyclin forms a complex with CDK2 and functions as a regulatory subunit of CDK2, whose activity is required for the G1/S transition of the cell cycle. This protein accumulates at the G1-S phase boundary and is degraded as cells enter S phase. Overexpression of this gene has been observed in many tumors, leading to chromosomal instability and potentially contributing to tumorigenesis. Diseases associated with CCNE1 include ovarian clear cell adenocarcinoma and retinoblastoma. Pathways associated with this gene include the cell cycle and the G1-G1/S phase of mitosis. Gene ontology annotations associated with this gene include protein kinase binding and kinase activity. An important homolog of this gene is CCNE2. | |||||||||||||||||||||||||||||||||
| 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, cell culture supernatants, and other biological fluids |
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4.1 ★★★★★
Based on 17 reviews
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Product Reviews
★★★★★ 4
Had no issue and it was very easy to use. It has been a week and no leak.
Size: 01- 24pcs (Classic Green)
Easy to use and its been a week and no leak. The kit is great and easy to use. The hard part was for me to get the screw out of my tire because the head was half gone. Once I got the screw out I followed the instructions and reamed the hole and inserted the plug and trimmed the plug.
If you plan on keeping it in your car you probably should add a screwdriver and/or pliers to get the screw out of the tire and a razor to trim the plug once it is in. You really need a razor to trim the plug I tried with a knife but the had to get a razor.
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Reviewed in the United States on April 5, 2025
★★★★★ 5
Great Item
Size: 01- 24pcs (Classic Green)
Nice kit. Has athe tools and pieces for many easy repairs. Fits neatly inside durable case.
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Reviewed in the United States on May 25, 2026
★★★★★ 5
Quality product
Size: 07- 22pcs (Blue)
Great kit for the price. Reamer and plug tool are strong and durable. Removable handles makes it great for broken tool or putting reamer on a drill if your lazy like me. Easy to use, easy to store and quality plugs
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Reviewed in the United States on November 30, 2025
★★★★★ 5
Emergency flat tire repair kit
Size: 02- 24pcs (Black)
Experience confidence on every road with the 24 Pcs Professional Heavy Duty Universal Emergency Flat Tire Repair Kit, the ultimate solution for fast, reliable, and long-lasting tire repairs. Designed for cars, motorcycles, trucks, trailers, RVs, ATVs, UTVs, and more, this kit provides everything you need to fix punctures quickly—without calling roadside assistance or waiting for a tow truck.
Built with high-quality, heavy-duty materials, each tool in the set is engineered for durability and performance. The solid steel T-handle reamer and inserter tools offer maximum strength, giving you the leverage and control needed to remove debris and plug punctures with ease. The professional-grade rubber repair plugs are strong, flexible, and designed to create an airtight seal, ensuring your tire holds pressure even in demanding conditions. Whether you’re dealing with a nail, screw, or sharp stone puncture, this kit helps you handle the situation safely and efficiently.
The set includes 24 essential pieces, such as repair plugs, valve stems, valve caps, a pressure gauge, lubricant, and additional accessories that make the repair process smooth and straightforward. Every item fits neatly into the compact, shock-resistant storage case, making it easy to carry in your vehicle’s trunk, glove box, or toolbox. This kit is perfect for emergency situations, long road trips, off-road adventures, work vehicles, and everyday drivers who want to be prepared for anything.
Even if you’re not a mechanic, the repair process is simple. The kit includes all the tools needed to clean, prepare, and seal the puncture, allowing you to get back on the road in minutes. It’s a cost-saving, time-saving, and stress-reducing solution every driver needs.
Stay prepared, stay protected, and stay moving with the 24 Pcs Universal Heavy Duty Tire Repair Kit—a reliable companion designed to handle unexpected flats with professional power and everyday convenience.
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Reviewed in the United States on November 25, 2025
★★★★★ 5
Convenient
Size: 01- 24pcs (Classic Green)
Very nice kit with everything in one zippered pouch.
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Reviewed in the United States on May 28, 2026