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Description
Rat MSLN 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. 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. 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. 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 mesothelin (MSLN) 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 mesothelin (MSLN) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Mesothelin | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Mesothelin, also known as MSLN, is a protein encoded by the MSLN gene. It is a 40 kDa protein expressed in mesothelial cells. The protein was first identified by its reactivity with the monoclonal antibody K1. Subsequent cloning studies revealed that the gene encodes a precursor protein that is processed to produce mesothelin, which is attached to the cell membrane via a glycophosphatidylinositol bond and a 31 kDa shed fragment called megakaryocyte potentiating factor (MPF). Although it has been proposed that it may be involved in cell adhesion, its biological function remains unclear. Knockout rat strains lacking mesothelin reproduce and develop normally. It is overexpressed in several human tumors, including mesothelioma, ovarian cancer, pancreatic adenocarcinoma, lung adenocarcinoma, and bile duct carcinoma. It binds to MUC16 (also known as CA125), suggesting that its interaction with MUC16 may contribute to tumor implantation and peritoneal dissemination through cell adhesion. A 64-amino acid region (residues 296–359) at the N-terminus of cell surface endothelin has been identified as the functional binding domain of MUC16 (termed IAB), suggesting a mechanism by which it acts as a functional chaperone of MUC16 in 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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4.3 ★★★★★
Based on 1114 reviews
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Product Reviews
★★★★★ 5
Great for mental stimulation. Durable for my chewers!
Color: Purple Lil Snoop, Color: Purple Lil Snoop
Love these! Quality material, safe for dogs — I make my dogs home made freezer treats and fill these up with it, and serve. Keeps them busy for hours. It’s also fun for them to roll it around to get regular treats out of. Toy + treats = a very happy pup. This size I got for my puppy. The larger ones I use for my adult dogs. Super cute too, and silent! I have Mini Aussies, so it’s great mental stimulation for them. Tires them out.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 7, 2026
★★★★★ 5
Very durable
Color: Blue
I have never written a review but just have to say how much our Pitbull LOVES this toy. It is very sturdy very good since he is such an aggressive chewer. This is only the second product I have found that he can’t destroy in 5 minutes. It keeps him busy for a long time and plays with it long after the treats are gone which is good as too many treats and he would gain weight. Highly recommend it. He weighes 70 lbs and the largest one is the perfect size. Kibble doesn’t work as it empties out immediately. I use Beggin Strips cut to size just to fit in the hole—they will come out but not all at once and survive his chewing on the toy.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 8, 2026
★★★★★ 4
Overall good toy for dogs
Color: Blue
My dog loves her snoop. I put little training treats in there for her, and she loves to roll it around and get the treats out. For her, it doesn’t last very long because they do fall out pretty easy, but she is a big dog so a smaller dog might have a harder time than her. Very sturdy and easy to travel with. Not noisy at all. I would say it’s a good, quick play toy for a dog like mine.
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Reviewed in the United States on April 14, 2026
★★★★★ 5
Fav Chew Toy 🦴
Color: Blue, Color: Blue
My baby is half rottweiler and half doberman &’ he has a pretty strong grip when it comes to chew toys. But this one, it’s his favorite toy! He never leaves the house without it. It’s also really fun to add his favorite treats in there when’s he bored to keep him entertained! Not loud at all and will keep him from chewing on other things and not heavy as other rough chew toys!
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Reviewed in the United States on May 31, 2026
★★★★★ 5
Great you made super well
Color: Blue Lil Snoop, Color: Blue Lil Snoop
This is such an awesome toy. It’s super durable strong bounces and a perfect size. We got the small. We got blue because our pup can see it. It allows for creativity and endless limits to what you can make for it to be a fun experience. It has a nice sized hole in the center which allows you to add treats and food. Pop it inward and it slightly if large enough prevents the treats from coming out super fast. The ball rolls bounces and they can grip it nicely.
It is easy to clean
The only thing I didn’t like was the way the plastic tabs were situated inside the toy as you just be careful you don’t allow it to drip inside and dog gets it. No noise just fun. Our 15lb Cavapoo loves it
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Reviewed in the United States on February 17, 2026
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