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Description
Rat FAK 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 PTK2 protein tyrosine kinase 2 (FAK) 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 PTK2 protein tyrosine kinase 2 (FAK) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat PTK2 protein tyrosine kinase 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Focal adhesion kinase (FAK), also known as PTK2 protein tyrosine kinase 2 (PTK2), is a protein encoded by the PTK2 gene. Most cells, with the exception of certain blood cell types, express FAK. Its activity triggers intracellular signaling pathways that promote the turnover of cell-cell contacts and the extracellular matrix, thereby promoting cell migration. FAK is required during development, and its loss leads to lethality. This 125 kDa protein is recruited to participate in the dynamics of focal adhesions between cells and plays a role in motility and cell survival. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.6 ★★★★★
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Product Reviews
★★★★★ 1
Received Burst Water Flosser, that never worked. Found out too late.
Color: Rose Gold
Ordered this item on recommendation of my dental hygenist, then needed a tooth pulled and was starting the dental implant process so was told I couldn't use it for awhile. The process took another 1 & 1/2 years. After the final crown I was told to use a waterpik. I went to use this, was unable to charge it, may have been missing a part, tried a matching plug. It wouldn't charge or turn on. Too late to return, so I'm out a good chunk of money in exchange for junk, something that never worked. I wonder if someone had returned a defective one as it had a dark finger smudge where it turns on, which I couldn't wash off. The tips were in a new package. Very disappointed.
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Reviewed in the United States on May 16, 2026
★★★★★ 5
Perfect in every way!
Color: Rose Gold
This flosser is perfection. It starts with the packaging and presentation. The flosser itself is beautiful (I got the rose gold), a nice sleek design that looks great on the bathroom counter. It is well constructed; I have tried the cheaper versions of these flossers and they all have a hose that hangs in the water reservoir that eventually gets clogged and stops working. Save your money and go for this one. Another great feature is how LONG it holds the charge. I am one month in and it is still operating on the initial charge at full strength. This makes it ideal for travel, along with the adorable travel bag included in the package. I also appreciate not having the wires cluttering up my counter space, which was the case with the cheaper models that have to be charged after each use. So far the company has been great to deal with - it comes with a 1 year warranty, which is easy to sign up for on their website. After the one year, you can subscribe to automatic tip replacements (each 6 months) and that will extend the warranty.
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Reviewed in the United States on March 21, 2023
★★★★★ 5
Be sure to still floss
Color: Rose Gold
This is a great product. However, make sure you still floss. I used the water flosser only between my dental cleanings and I ended up with a lot of tarter that had to be scraped off. I was told that I should continue using string floss as plaque is sticky and a water flosser can leave some behind. The hygienist compared it to expecting rain clean your car well. I still give it 5 stars as it remove particles from between my teeth that floss leaves behind. Also, my teeth were nearly perfectly clean during my last visit to the dentist. There was virtually no tarter as I followed her recommendation to both string and water floss.
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Reviewed in the United States on January 5, 2025
★★★★★ 3
They leak and the don’t last, but they’re sleek.
Color: Black
I love the shape of this flosser, that aspect is perfect. That’s the reason I’ve bought (3) over the years. But unfortunately, I’m done. Looking for a new gig. Every single one has leaked water. I just dealt with it. But the year and a half life they last is the deal breaker. Buy the insurance. You will use it. They simply do not last forever.
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Reviewed in the United States on March 22, 2026
★★★★★ 5
Perfect for home or traveling
Color: White
This is my second water floss, product, and by far the best. Yes, the water reservoir is not as large as some of the others, but it is perfectly adequate. Easy to use, has three different settings, that are easy to change. Definitely enough power to get all the food out from between my teeth. I use this when I had braces, and I use it now with my retainer, so that my teeth are always clean. I like the design, and the fact that it doesn’t have a tube to suck the water. The charge lasts for an extended period of time and recharging is very quick. I can go two to three weeks, and it’s still got a good charge to it. Highly recommend.
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Reviewed in the United States on July 23, 2023