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Description
Human ISM1 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 an isthmin 1 (ISM1) 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 isthmin 1 (ISM1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Isthmin 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | ISM1 (Isthmin 1) is a protein-coding gene important in tumorigenesis and invasion, including in colorectal cancer (CRC). Conditions associated with ISM1 include introitus testicular hematoma. An important homologue of this gene is THBS4. | |||||||||||||||||||||||||||||||||
| 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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4.3 ★★★★★
Based on 26 reviews
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Product Reviews
★★★★★ 4
Well made and durable, not a toy the pups seek out
Size: 1 Pack, Style: Tug
Well made. Thought our staffies would love since they live tugging with each other. Sadly not something they seek out. We love ALL the benebone chews though!
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Reviewed in the United States on April 6, 2026
★★★★★ 5
Durable fun enrichment toy
Size: 1 Pack, Style: Pawbler
I had been wanting to buy one of these kinds of toys for some time but thought that the other brands I had seen were too pricey so this was a great compromise for the price. My 5 yr old Portuguese water dog loves it. He spent 25 min playing with it the first day. The trick is to find a treat that is just big enough where it doesn’t fall out right away so it makes it challenging and fun for them. He has been chewing on it a lot for a week and it has not broken or shown signs of damage. One thing to note is that there is a small hole on the bottom of the toy so you cannot freeze liquids in it without covering the hole in some way so that would be my only complaint. Besides that it’s a great toy that keeps him busy and happy.
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Reviewed in the United States on September 1, 2025
★★★★★ 3
Dog doesn’t care for it
Size: 1 Pack, Style: Bone
Hard to review product for its durability because my dog has never chewed it once. Doesn’t care about it at all. The bone feels rugged but smells like playdoh. It’s sat on the floor for over a month untouched.
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Reviewed in the United States on May 6, 2026
★★★★★ 1
Pawbler FAIL: First time our dog bit the ball, it popped open and dumped all the kibble.
Size: 1 Pack, Style: Pawbler
I would not recommend! We have a 48# dog and within minutes of giving it to him, the Pawbler was open and all of the kibble was in a pile on the ground. I had washed and dried the Pawbler, added kibble, and tightened the lid as tight as possible. Imagine my surprise when, within minutes of giving it to him, the lid was off and all the kibble was on the ground. I thought it was a fluke so then I made an extra effort to make sure the lid was super tight...and the same thing happened. This time there was no food inside so he started chewing on the two parts. Within minutes of that, he'd chewed/damaged the rubber on both the body and the lid. I took it away as I didn't want him to destroy/eat any of the rubber bits. I was surprised he could even get the Pawbler in his mouth. It's heavy and he is a medium sized dog. He has the Benebone Bone, the WestPaw bone, the WestPaw ring that he chews on all of the time and none of those are showing any type of wear so to see this opened/chewed within 15 minutes makes me think this was defective. I am returning and hoping for a refund. DO NOT BUY! Poor value for the money. Not durable. Not chew resistant.
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Reviewed in the United States on April 30, 2026
★★★★★ 5
Just what I was looking for to slow my large dog while eating
Size: 1 Pack, Style: Pawbler, Size: 1 Pack, Style: Pawbler
I needed something for my 50-pound dog to slow down her eating. The usual slow-feeder bowls and mats weren't really slowing her down much and I wanted her to have to think a little bit. She's not smart enough for puzzles though. So I got this kibble dispensing toy. My dog is not a chewer so I don't have to worry about her destroying it.
LOVE: holds a little more than a cup of kibble, dispenses different sized kibbles, heavy weight so it wobbled around a lot without going too far away, QUIET- I couldn't bear the noise of hard plastic dispensers clacking around and this one is very quiet on our hardwood floors, kept the dog busy for at least 20 minutes. It's really all I was hoping it would be. And, well, as you can see in the video, the cat is happy about it too...
As with all of these dispensers, there will be a couple of kibbles left inside that are hard to get out. But that gave my dog something to do overnight. She spent a long time getting that last kibble out. The slight scent they add to the natural rubber is not a problem at all. This dispenser is absolutely worth the money, it's going to last us a very, very long time.
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Reviewed in the United States on October 17, 2025