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Description
Rat CFL1 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 ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected 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 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 cofilin 1 (CFL1) capture antibody. After incubation and washing, the sample 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 cofilin 1 (CFL1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Cofilin 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Non-muscle cofilin-1 (CFL1) is a ubiquitously distributed actin regulatory protein that binds and depolymerizes filamentous F-actin in a pH-dependent manner and inhibits monomeric G-actin polymerization. It is involved in the transport of actin-fibronectin complexes from the cytoplasm to the nucleus. Reelin signaling leads to serine 3 phosphorylation of cofilin-1, and this interaction may play a role in the regulation of Reelin-related neuronal migration. It has been shown to interact with HSPH1 and LIMK1. | |||||||||||||||||||||||||||||||||
| 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 and other biological fluids |
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4.6 ★★★★★
Based on 13 reviews
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Product Reviews
★★★★★ 5
Favorite Toy
Green and brown dog toys, sturdy and safe. Dogs love chewing them, good for teeth cleaning.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 21, 2026
★★★★★ 5
Durable chew toy that keeps my puppy engaged
Size: X-Small, Color: Tangerine, Size: X-Small, Color: Tangerine
My dog went crazy for this toy the second I gave it to him.
He is an 8-month-old 7.5 pound Yorkie puppy, an aggressive chewer on the tail end of teething. He has shown no mercy to this toy and yet there’s not a dent in it. That’s why I got it because the material holds up to aggressive chewing. My puppy has confirmed that!
It keeps him entertained for a long time. It’s small enough to fit in his mouth for chewing but not so small that he would choke.
He loves to sit on the couch but is too young to jump off. This toy keeps him entertained on the couch for a long time so he stays put. He is totally focused on the toy. It is the perfect toy for a bored chewer. Works really well. And it’s super easy to clean.
Because it’s such a small toy, I’m only letting him use it when I’m nearby to supervise. But it has really held up to those busy teeth and Yorkie jaw.
It’s hard to find a durable toy that also holds his interest. This seems to check all the boxes. I didn’t even have to coat it with a treat taste to draw his interest.
I think the bone shape is very functional and attractive for him. He can go a solid 30 minutes chewing on it without getting bored or distracted, and that’s a long time for a Yorkie puppy.
I got this toy because I noticed he wanted to chew, but I was tired of other toys that came apart. This one seems like the right durable choice.
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Reviewed in the United States on January 17, 2026
★★★★★ 5
Best it for dogs that destroy all other toys
Size: Large, Color: Granny Smith
We bought his it for our dog in 2020 when we rescued her. She has literally destroyed every single toy she has within minutes of giving them to her, but not this toy. It has lasted almost 6 years. It finally started to tear in the center so we bought her another one. Honestly it probably broke from my kids playing tug of war with her and they toy and that’s not really what it’s designed for. I’d rate this a 10/10. It’s quality but no so hard she’ll hurt her teeth.
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Reviewed in the United States on April 17, 2026
★★★★★ 5
Finally an all-purpose toy that doesn’t fall apart!
I thought there wasn’t a toy in the world that could withstand the craziness of our 95 lb German Shepherd, until I found this! It’s great for rolling, tugging, fetching, and chewing, soft enough rubber for his teeth yet so far solid enough to not fall apart. He’s had it for almost a month and I don’t see any rips, holes, or missing pieces. I don’t know if it floats since we use it in the house. And a HUGE bonus: IT DOESN’T SQUEAK. 🥳🥳🥳 We ordered a size Large and it’s a perfect fit. He loves to grab and carry it around when he’s excited, so it’s solid but not too heavy. Will definitely be checking out more toys from this brand. Thank you West Paw!
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Reviewed in the United States on April 15, 2026
★★★★★ 4
Durable with limitation
Size: Large, Color: Granny Smith
offers good durability, but our Olde English Bulldogge was able chew an end section nearly off so cut it off and allow him to enjoy the Zogoflex under supervision
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Reviewed in the United States on February 24, 2026