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Description
Rat TJP1 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 capture antibody against Tight Junction Protein 1 (TJP1). 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 Tight Junction Protein 1 (TJP1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Tight Junction Protein 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Claudin 1, also known as ZO-1 or TJP1, is a peripheral membrane protein encoded by the TJP1 gene. It belongs to the zonula occludin family (ZO-1, ZO-2, and ZO-3), which are tight junction-associated proteins, of which ZO-1 was the first to be cloned. It was first isolated in 1986 by Stevenson and Goodenough using a monoclonal antibody raised in rodent liver to identify a 225-kD polypeptide in whole liver homogenates and tight junction-enriched membrane fractions. It functions as a scaffolding protein, cross-linking and anchoring tight junction (TJ) tethers, fibril-like structures within the lipid bilayer that connect to the actin cytoskeleton. The encoded protein may be involved in signal transduction at cell-cell junctions. Two transcript variants encoding distinct isoforms have been identified for this gene. | |||||||||||||||||||||||||||||||||
| 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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4.6 ★★★★★
Based on 28 reviews
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Product Reviews
★★★★★ 5
Surprisingly Tough Chew Toy for Aggressive Chewers
Size: Medium & Large, Color: Red Scorpion, Size: Medium & Large, Color: Red Scorpion
This chew toy is made from a very hard nylon style material and feels substantially tougher than the average dog toy. It definitely seems geared toward aggressive chewers and larger dogs that destroy softer toys quickly. Our pup immediately liked it, especially with the peanut butter groove feature, which helped keep them interested longer.
The toy feels almost indestructible during normal chewing sessions and has held up very well so far with no major gouges, cracks, or chunks coming off. Based on first impressions, I would expect this to last at least a month or more for many dogs, which is far better than a lot of chew toys in this price range. Another nice bonus is that it is dishwasher safe, making cleanup easy after using peanut butter or other treats in the groove.
As for the teeth cleaning claim, I honestly cannot comment much on that yet. I have not personally noticed any obvious difference in our dog’s teeth or dental condition from using it. Still, as a durable boredom relief chew toy for aggressive chewers, it has been a solid performer so far and seems like a good value.
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Reviewed in the United States on May 27, 2026
★★★★★ 5
My dog approves
Size: Medium & Large, Color: Red Scorpion, Size: Medium & Large, Color: Red Scorpion
My dog absolutely loves these flavored chew bones! She’s a very aggressive chewer, and this toy has held up really well to her constant chewing. It keeps her entertained for a long time and seems to be one of her favorite toys.
I also love the unique scorpion shape. Living in the desert, we see scorpions pretty often, so I thought it was a funny and fitting design. While I don’t expect it to turn her into a scorpion hunter, it definitely adds a fun touch!
The flavor keeps her interested, and the durable material makes it a great option for heavy chewers. Overall, a great toy that has been a huge hit with my pup. Will definitely be purchasing more!
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Reviewed in the United States on May 29, 2026
★★★★★ 5
Cheaper than shoes!
Size: Medium & Large, Color: Red Scorpion, Size: Medium & Large, Color: Red Scorpion
Instant hit with a dog that likes chewing shoes! This is a much cheaper alternative than a new pair of shoes. They state it is indestructible but at the same time the directions state when it reaches a certain chewed up point to take it away. So obviously this is destructible but it does allow for a lot of chewing. I can not personally attest to the flavor but it must be good because the dog wanted this chew before i even unwrapped it. It is somewhat ugly in both color and shape but I have had no complaints from the dog! I also appreciate that this can be put in the dishwasher to clean. To be honest i have never considered that with other toys and I do appreciate that it can be placed in the dishwasher. Most importantly, this lasts, the dog is in love with this, there are multiple limbs for the dog to chew. It’s his new favorite toy and worth every bit of the cost!
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Reviewed in the United States on June 3, 2026
★★★★★ 5
Durable
Size: Medium & Large, Color: Red Scorpion
This bone has proved to be durable and tasty (per our Labrador). She chews and chews with barely a dent. I’m guessing the flavor is good because it’s still her go-to bone after almost a week. Beware of your pet dropping it on your toes though as it is quite hard.
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Reviewed in the United States on June 2, 2026
★★★★★ 5
Well-made and great size for the price!
Color: blue black
Very tough product, no odor of plastic, good toy for outside, perfect for medium or large dogs. Noise isn't too bad and the color is cute as well. Can't destroy this toy!
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Reviewed in the United States on May 26, 2026