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Description
Rat PER2 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant and centrifuge at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. Cell Supernatant: 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., 1 g of tissue sample to 9 mL 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 the tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000 × g for 5-10 minutes, and the supernatant can be assayed. Preparation before testing: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the gradient working solution of the standard: 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 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 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 Period Circadian Protein 2 (PER2) capture antibody. After incubation and washing, the assay 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 Period Circadian Protein 2 (PER2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Period Circadian Protein 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | PER2 is a protein encoded by the PER2 gene in mammals. PER2 has attracted attention for its central role in circadian rhythms. Disorders associated with PER2 include advanced sleep phase syndrome, familial 1, and advanced sleep phase syndrome. Pathways associated with PER2 include the metabolism and effects of the circadian clock and melatonin. Gene ontology annotations associated with this gene include transcription factor binding and transcription cofactor activity. An important homologue of this gene is PER1. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, cell supernatant, tissue homogenate, etc. |
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4.5 ★★★★★
Based on 431 reviews
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Product Reviews
★★★★★ 5
Durable , dogs love them !
Size: 2 Count (Pack of 1), Style: Pack of 2, Size: 2 Count (Pack of 1), Style: Pack of 2
These chuck it balls are a fan favorite of my own dogs and my foster pups . So much so I have sent my foster home with them when they go to their new families! I seriously need an endless supply !
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Reviewed in the United States on April 30, 2026
★★★★★ 5
Whistles when thrown
Size: 2 Count (Pack of 1), Style: Pack of 2
One of my shepherd’s favorites! Truly does whistle when you throw it! Good quality!
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Reviewed in the United States on May 25, 2026
★★★★★ 5
Whistle away my friend!
Size: 2 Count (Pack of 1), Style: Pack of 2
Love it! My dog loves it! The people at the dog park love it! The whistle is the best thing about these balls but the hard durable rubber and the denseness of the ball really make it fly. Let me quickly breakdown each of these:
First the sound is like a...well, whistle coming from a human but oscillating and slightly different. It helps the dogs track where it goes and helps them find it better when it is getting dark (while flying of course, not when it is stationary). It barely makes a sound when throwing it with the wind but really howls when throwing it into the wind. This is why you get this ball and most everyone at the dog park perks up and wonders what that is, asks about it, and thinks it is very cool; which it is!
Next is the durable rubber which doesn't have a single bite mark or split yet in 4 months of constant daily play for about 1 1/2 hours every day! The dog chews it as he brings it back and other dogs steal the ball and run with it while "killing" it perpetually and it has held up great. I don't claim this is going to be Kong ball durable but it is very well chosen rubber.
Last is the denseness of the ball which really makes to go far in a chuckit launcher. One doesn't really notice how far this goes until going back to a tennis ball and realizing how short a tennis ball goes compared to this. The whistler ball, and presumably other chuckit rubber balls (glow, irregular bounce), go 30-40% farther when I throw then using the longest chuckit thrower. This is excellent and wears my retriever out even faster; not to mention the rippling muscles he has from sprinting that whole way!
Overall I got both ball several months ago and we started with the blue ball and haven't had to touch the orange one yet. This is his goto ball at the park and it is the most popular ball, bar none, for all the dogs to steal. The bright blue aspect, the rubber aspect, the sound, and exclusivity of it make it coveted by dogs and owners. I might be gushing and glowing over this ball a bit much but I tend to do that when something unexpected ends up being part of your everyday life and blows you away at how wonderful it is.
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Reviewed in the United States on December 20, 2012
★★★★★ 5
Great for dogs that are chewers
Size: 2 Count (Pack of 1), Style: Pack of 2
These are wonderful for our dog. The bright colors help us find them when our dog can't. The size is perfect to play fetch. Our dog will chew on it a bit and it stays together and does not fall apart. That is a big bonus because our dog is a chewer!
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Reviewed in the United States on February 4, 2026
★★★★★ 5
It's A Whistler!
Size: Medium (Pack of 1), Style: Pack of 1
This Chuckit! Whistler Ball is basically a regular fetch toy that decided to become an annoying little teakettle mid-air. My dog goes absolutely nuts for it—every throw sounds like a tiny referee is shrieking "foul!" the whole way across the yard. He chases it harder than he chases the mailman, and bonus: I can actually find it in tall grass because it's screaming for help. Super bouncy, tough enough that he hasn't destroyed it yet (miracle), and now our walks sound like a low-budget horror movie.
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Reviewed in the United States on February 14, 2026