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Description
Rat ARNTL ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and pipette 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: Tissue homogenates and other biological fluids Preparation before testing: 1. Please take out the test kit from the refrigerator 10 minutes in advance and
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and pipette 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: Tissue homogenates and other biological fluids Preparation before testing: 1. Please take out the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare a gradient standard working solution: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 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 500 μL of universal diluent to each tube. Pipette 500 μL of the 10 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5 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 an aryl hydrocarbon receptor nuclear translocator-like protein 1 (ARNTL) 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 aryl hydrocarbon receptor nuclear translocator-like protein 1 (ARNTL) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Aryl hydrocarbon receptor nuclear translocator-like protein 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Aryl hydrocarbon receptor nuclear translocator-like protein, also known as ARNTL or brain and muscle ARNT-like protein 1 (BMAL1), is a protein encoded by the BMAL1 gene. BMAL1 encodes a transcription factor with a basic helix-loop-helix (bHLH) and two PAS domains. This 62-amino acid protein plays a key role as one of the positive elements in the mammalian autoregulatory transcription-translation negative feedback loop (TTFL), which is responsible for generating molecular circadian rhythms. | |||||||||||||||||||||||||||||||||
| 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 | Tissuehomogenatesandotherbiologicalfluids |
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4.0 ★★★★★
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★★★★★ 5
Little Small On The Wrist
Really cool timepiece, been loving timex a lot recently, this is me coming over Casio/G-shocks. Although it is a bit smaller on the wrist it still looks very nice and I really love the gray and like green color contrast. Really easy to read the time as well.
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Reviewed in the United States on May 27, 2024
★★★★★ 1
Sturdy and digital
This watch has “ fake screws” holding the band on . The band is actually held on by the usual pins holding it on most watches but weaker . The pin came off while I was putting it on for the first time! This is the second time I bought off Amazon in 2 weeks where the watch appeared to be sturdy and the pins fell off . Going back to military watches that have material straps that go completely around wrist and do not have pressure on pins
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Reviewed in the United States on May 8, 2026
★★★★★ 5
Ten years of ownership…Thank you, Timex!
Color: Black, Size: One Size
Solid watch for everyday wear. I have worn this watch for over 10 years. I let a watch shop replace the battery about every 18 months. Other than the original band (which lasted 5 years) I have had zero problems. Still looks great, rugged and manly😇
Overall I recommend this watch for its value, reliability, simple clear design, legible hands in all light conditions. Great watch!!!
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Reviewed in the United States on February 13, 2026
★★★★★ 5
Solid Watch for Everyday Casual Wear
Color: Black, Size: One Size
I have purchased this item other times and I’m quite satisfied with wearing this watch everyday with casual wear. The size is just right not too small or large.
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Reviewed in the United States on May 23, 2026
★★★★★ 4
Nice simple effective inexpensive watch
Color: Black, Size: One Size
I just ordered my second one of these. They are cheap, keep good time, show me the date, and last a long time. My last one lasted over 10 years. The only thing wrong with the last one was that the battery needed replacement (I think it was only the second time it had needed a battery in all the years I had owned it) at the same time the band wore out and broke. Rather than buy a new battery plus a new band, I just got a new one. I am hard on watches, and this one stood up very well. I regularly shower and swim with it on, and no problems.
It doesn't have a lot of functions I don't need, like stopwatch, multiple time zones, etc. I have found those things more of a bother than a benefit, because any time you bump the knob against something, the display changes and you're always pressing all the buttons multiple times just to get the simple info needed- like what time is it and what is the date
DEC 2020 UPDATE - I am ordering another one of these. The last one, I bought in mid 2019. Early in 2020, the date function quit working. By the end of November, the whole thing quit working. I know it's not the battery, because the light still works. So I am reducing my review by one star, and we'll see how well a new one does. The watch before the current one lasted about 10 years, so getting only a little over a year out of the current one is not good. Perhaps the current one just wasn't as good as the previous ones. It's entirely possible that they keep making the same watch out of lesser and lesser materials and workmanship.
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Reviewed in the United States on November 21, 2016