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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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4.3 ★★★★★
Based on 22 reviews
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Product Reviews
★★★★★ 5
Light weight & Sturdy
Size: 6 Panel, Color: Dark Coffee
I am so pleased with them. My lanai gets morning sun and I am concerned about the UV rays fading the
fabric of my furniture No more having to cover each piece at night. The screens are great.
Thank YOU
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 10, 2025
★★★★★ 5
Good
Size: 6 Panel, Color: Dark Coffee
Good, no complaints after several months of use
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Reviewed in the United States on January 15, 2026
★★★★★ 3
Functional divider for windows privacy
Size: 8 Panel, Color: White, Size: 8 Panel, Color: White
I ordered both a six and eight panel version of the white divider. There was some confusion on the FedEx deliveries. The eight panel divider arrived at first, but Amazon showed both as delivered. I had to contact both the seller and Amazon to learn that the six panel divider was to arrive the following day by FedEx, which did occur. Both the boxes were a bit dirty and had some small punctures. This divider had a top area and bottom area set of plastic wrap tapes, both of which were loose or undone. (The eight panel divider was not so, and properly wrapped.) The top of thid divider looked a little dirty, but passable/cleanable. The seller should consider wrapping the entire product with bubble wrap to protect against potential delivery/shipping physical and water damage. (FedEx left the box by my outside door, which could have gotten potentially soaked in rain. One better be home to take delivery if the weather is bad.) This said, the divider was easy to set up, with all the metal hinges working. No assembly was required, just unboxing and standing up. The left side was a little uneven on the bottom with the first panel, but ok. In combination with my mini-blinds, it provides the additional layer of privacy I desired. I even don't notice outside headlights and tailights through the windows with my room lights on. Additionally, the divider is easy to adjust to allow access to my windows for opening and closing. So, no need for me to hang curtains and damage my drywall. Overall I give the product a C grade because of the lack of protection for the FedEx shipping. (As a former University told me years ago, "The A has been cheapened. In this class you will work for an A." And, I got a B. So, I am a bit jaded. :-)) Lastly, if one had cats and dogs, or even small children, the divider could be easy for them to topple over. I would not consider these dividerd in that case. My motto is, "Safety first."
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Reviewed in the United States on December 31, 2025
★★★★★ 5
Prrfect
Size: 6 Panel, Color: Dark Coffee
Love it. Perfect for my needs to create a hidden storage space in my spare room. Highly recommended
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Reviewed in the United States on January 21, 2026
★★★★★ 5
Nice quality
Color: Rustic Brown
Cute and nice quality.
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Reviewed in the United States on May 9, 2026
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