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Description
Rat ECM1 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 specimens using EDTA or heparin as an anticoagulant and centrifuge at 1000×g for 15 minutes at 2-8°C within 30 minutes of collection. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 3. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. Pre-Test Preparation: 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 stand for 15 minutes to completely dissolve, then gently mix (concentration is 3000 pg/mL). Then dilute to the following concentrations: 3000 pg/mL, 1500 pg/mL, 750 pg/mL, 375 pg/mL, 187.5 pg/mL, 93.75 pg/mL, 46.875 pg/mL, and 0 pg/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 3000 pg/mL standard working solution into the first EP tube and mix thoroughly to make a 1500 pg/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 extracellular matrix protein 1 (ECM1) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of extracellular matrix protein 1 (ECM1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Extracellular Matrix Protein 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Extracellular matrix protein 1 is a protein encoded by the ECM1 gene. The extracellular protein encoded by this gene contains a cysteine pattern characteristic of the ligand-binding "double loop" domain of the albumin family. This gene maps outside the epidermal differentiation complex (EDC), a cluster of three gene families involved in epidermal differentiation. | |||||||||||||||||||||||||||||||||
| 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 | 46.8-3000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, cell culture supernatant |
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4.1 ★★★★★
Based on 5 reviews
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Product Reviews
★★★★★ 5
Super fun for little dogs
Color: Burger Shape Assorted Color, Size: Small
These are so fun! I have two small dogs (10 lbs) and they both love them:) good squeak and just the right size for them. They get a little crazy with them but they love it!
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Reviewed in the United States on April 25, 2026
★★★★★ 5
Only One of Two Toys That Any of My Three Dogs Will Play With
Color: Burger Shape Assorted Color, Size: Small
I can’t overemphasize this enough:
My pooches just aren’t keen on toys. I don’t know why. I’ve tried everything to get them to play with SOMETHING, instead of just lying around, or begging ME to do all the work to entertain them all the time. But whenever I buy or make one for them, all three just look at me like, “Meh. Bored. Try again, Ma.”
Until I bought these. I’ll spare the reader a lot of details, but for whatever reason, these little, round, mouth-sized squeakers were a hit. There’s literally nothing special about their construction or functionality at all, but my dogs think they’re great. Kinda like how kids don’t care about the fancy, expensive toy you bought them, but prefer to play with the box it came in. 🤷♀️ I was concerned about them getting chewed up the first day (or worse, getting swallowed), or me just being driven insane with the squeaking, but only the latter has been an issue, and only the first few days. My dogs are very diverse mixed breeds and sizes, and all three think these are the best thing ever. My medium-sized bulldog is so adorable: He very gently picks one up, squeaks it a little bit, and then runs around holding it in his mouth until he decides to drop it (usually 30 seconds or less). Then my big dog swooshes in and steals it from him (It’s a “game” that only one of them enjoys). The biggest annoyance with these is having to go pick them up wherever my poorly-trained doggos drop them (but that’s my fault, of course).
In a nutshell: They’re a hit at my house. Just be vigilant the first few days to make sure your dog isn’t interested in trying to eat them.
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Reviewed in the United States on July 27, 2023
★★★★★ 5
Hands down, the best dog toys on the Planet!
Color: Burger Shape Assorted Color, Size: Small
Our Miniature Pinscher absolutely loves squeak toys. We tend to get those long-bodied, no-stuffing animals which are great for tug-a-war, but on her own, she rips these suckers apart and removes the squeaker within a day of owning it. I occasionally buy a big bag of just squeakers and hide them in shoes and what not, but those have a 5 minute life span as she'll pull the little whistle right out.
These latex squeak toys are a total blessing. They're super well made and just the perfect dog toy. We've had them a week, and she's been playing with two out of the package and not a tear or rip. The squeaker is very loud and the material is impossible to rip. She's getting tons of exercise running around the house with these things, and loves when I throw 'em. Love the colors and faces on the burger style. Easily the best dog toy on Amazon and we've been through 5 years worth of purchases.
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Reviewed in the United States on August 30, 2024
★★★★★ 5
Great ball for small dog (3.5 pounds)
Color: Burger Shape Assorted Color, Size: Small
I have been looking for a ball that my 3.5 pound Maltese puppy can enjoy. Up until now it was small balls were difficult for my little Rosie to grab. These balls at first seemed to be too big. But after a few minutes Rosie grabbed one of the balls in her mouth and has been crazy for these balls ever since. They collapse so she can get her mouth on it. They squeak each time she bites down. When I toss the ball, they bounce around like crazy and make a squeaking sound. They are so soft that it would be difficult to break anything. Also since they are not round, they do not roll under furniture most of the time. I uploaded a video of my little Rosie fetching one of these balls. Great toy!
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Reviewed in the United States on May 11, 2022
★★★★★ 4
Dog doesn't chase balls but does chase these!
Color: Regular Ball Multi-color, Size: Medium
This is a great product. My Aussie mix won't chase regular balls, but will chase these for long periods of time. Its the only way I can get him excercise right now. He holds them in his mouth constantly and has lost interest in his kong stuffed toys. This is my second time buying. The first time was great but the second batch had hardly any sqeak and not much air. Seller- please check your quality control! I have to go to another seller if these aren't consistent, because my dog needs the squeak in order to be interested.
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Reviewed in the United States on March 26, 2025