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Description
Human ARL1 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: 1. 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: 1. 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. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen 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. 3. 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 the tissue and mince it. 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 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. 4. 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 Aldo-keto reductase family 1 member B10 (ARL1). 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 Aldo-keto reductase family 1 member B10 (ARL1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Aldo-keto reductase family 1 member B10 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | AKR1B10, or aldehyde-keto reductase family 1 member B10 (AKR1B10), is an enzyme encoded by the AKR1B10 gene. This gene encodes a member of the aldehyde/ketone reductase superfamily, which comprises over 40 known enzymes and proteins. This member efficiently reduces fatty and aromatic aldehydes, with less activity towards hexoses. It is highly expressed in the adrenal glands, small intestine, and colon, and may play a significant role in liver cancer development. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenates and other biological fluids |
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4.0 ★★★★★
Based on 17 reviews
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Product Reviews
★★★★★ 5
Great
Size: Medium, Number of Items: 1
I need something that was soft for my three year old and this ball is super great, it not only is the right size but it also fits in his mouth just right. Although he is a super aggressive chewer, the ball toy hasn't been totally destroyed yet and it is easy on his gums. The ball is so durable that got two of these toys just for fun.
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Reviewed in the United States on May 10, 2026
★★★★★ 5
The best!
UPDATE: 08/05/2025 - This is still one of my dog's 2 fav training/play toys. A couple of notes:
1. This is not a chew toy. If your pup is chewing up the rope, that's on you. That being said, you can get 100' of the poly cord rope at Home Depot for like $4.98. You can make nearly 100 ropes out of 100'. How hard is it to replace the rope?????
2. My pup is a now 15 month old Malinois. His original balls, purchase in September 2024 are still in everyday, multiple times per day, play. I did get careless and leave one where he got it and chew the rope. Easily replaced. Nothing to fret over. Yes, that was on me!
3.The foam ball is virtually indestructible! Replace the rope!
4. I noticed the price for the yellow large went from $12 each to $24. THIS will cause me to go elsewhere if they doubled in price. I LOVE these balls but not that much. I noticed they have a new yellow ball that has a strap vs. a rope. I might try that if I need one, but I have 6 of these balls, 2 never used yet, 2 that are virtually new looking, and 2 that have been in play since september 2024. I don't think I'll be buying more any time soon, but if I do need more, I hope they haven't gone up to $24 each.
One of my top, go-to training tools. Quality is great other than the shrink wrap around the string joint. It's junk but not essential.
Great tug and fetch toy. Essential to teaching a good "out" command (2 are recommended for this).
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Reviewed in the United States on January 11, 2025
★★★★★ 5
Best dog ball
This is our go to dog toy for our German shepherd. its great for training in sports as well as outings at the beach, considering it floats! Never had him destroy one, so its extremely durable. I like that its a bright color so its easier to locate when i accidently let go of the rope too late and end up whipping it 20 ft into the woods. Easy to clean, all around just a great toy. Also love the large size as having a dog accidently lodge a ball into its throat is a real fear of mine, I do not have to worry about that with this toy. I should also mention the rope is a must, as touching a slobbery ball isn't the greatest feeling in the world and it puts your hands out of harms way. I will forever order these.
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Reviewed in the United States on September 7, 2025
★★★★★ 4
Great ball, bad rope
Size: Medium, Number of Items: 1, Size: Medium, Number of Items: 1
My dog and I love the starmark ball on a rope tug toys. While this is one of the best tug toys we have found, I am continually disappointed with the quality of the rope. The plastic sleeve comes off the first day, and after that the stitching begins to weaken until the rope ends come apart and the rope slides out of the ball. Is there a way to improve this design so that this toy lasts longer? My dog is not left alone with the toy, it is only used as an interactive tug toy as a training reward.
I am updating my review now several months later. I reinforced the rope with a paracord braid and the toy now lasts a very long time. My dog and I play with this toy everyday on our morning hike. He is a large German Shepherd with high ball drive. He carries this ball in his mouth several miles on our morning hike. We play fetch in the fields near our home and in the lake beyond the fields with this toy every day. This toy is his reward in obedience training and we play a lot of tug with it every day. Reinforcing the handle has made a huge difference as the toy lasts and lasts now with the improved rope. Great ball, we will always have a collection of them at our house!
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Reviewed in the United States on September 19, 2020
★★★★★ 5
Great for training and play!
Size: Medium, Number of Items: 1
My German shepherd love this toy! Great to take on walks with you as light weight and can fit in your pocket. Stands up to the toughest of play. Great as reward toy for training in place of treats! We always have one on hand at home or out and about!
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Reviewed in the United States on March 18, 2026