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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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4.5 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 5
Great product!
Color: Space Gray, Size: mStand, Style: mStand
I bought this laptop stand because I am was getting tired of looking down at my laptop screen, it was hurting my neck. Now it actually has helped me create a better setup, because my laptop is elevated and when I plug in my external monitor I can use the laptop screen at the same time, increasing my desktop space and increasing productivity since the screens are much closer to the same height now. I also like the cord management, it helps keep things cleaner. The space underneath works great to hold other things like my ipad when not in use, so my deskptop is less cluttered. I bought the one that does not swivel, its pretty sturdy and solid.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 28, 2020
★★★★★ 4
Fantastic stand that is great for posture. Nicely compliments the premium look of MacBooks
Color: Space Gray, Size: mStand, Style: mStand
I thought I wouldn't like this stand but I decided to purchase it anyways owing to the thousands of 5-star customer reviews.
I ended up loving it. It is perfect for a tall guy like me that needs to have better posture.
The rubber pieces on the surface not only prevent my MacBook Pro 15" from sliding, but also stop the stand from scratching the base of the Mac. The Space Gray color matches my MacBook Pro perfectly.
I thought this product was going to be way too tall because of the pictures but it ended up being just about right. If you don't like the height you can also go buy the
which is by the same company but is adjustable.
I love using the hole in the back for cable management.
It is also more portable than one might think. Yes, it doesn't collapse or fold or anything however, you can put it in a bag and it will leave a nice cavity for you to place books and other materials in. I have brought it with me to coffee shops.
The only reason I am giving this product 4-stars is that it is not perfectly stable. There is a slight wobble during typing. It is not extreme enough to really get in the way of use but it certainly isn't a perfect product like some people claim. Perhaps they were using lighter or smaller laptops.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 9, 2017
★★★★★ 5
I'll stick to Rain Design
Color: Space Gray, Size: mStand, Style: mStand
There are a lot of laptop stands out there and call me a loyalist since I bought my first Rain mStand over 10 years ago but I came back to Rain for my second one after giving the original one to my kid for college. I purchased the Space Gray version to match my current Macbook Pro color. It's still a handsome, well-crafted non-nonsense design. It's great that Rain didn't change the design much from the original one I had. It works well with padding in just the right places, and it's timeless design is a complement to any room decor. Go with Rain.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 29, 2024
★★★★★ 5
Simple, hefty configurable small tray
Color: Black, Color: Black
"Hefty" is not an expected description for a drawer organizer. But it is in this case because of the solid metal plate it's made from. You could brain a zombie with this tray and not dent it (the tray... the zombie's the one that would be dented). Guess the material choice does make sense since the two wooden dividers uses magnets to hold positions. A strong bump would be needed to shift the placed dividers.
You get some basic configuration choices from the dividers. Including additional (different length) dividers would allow further sectioning options would have been nice.
It also looks good too.
Like the design and utility. Would also go for a bigger version. Perhaps that one, like this, would have secondary uses... perhaps as armor plating?
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 19, 2026
★★★★★ 5
Elegant and easy to customize to organize your desk
Color: Black, Color: Black
This desk organizer was higher quality and more elegant looking than I thought it would be, so I decided to use it to hold some things on my dresser. It’s too pretty to hide away in my drawer.
The compartments snap into place with magents so you can adjust based on how you want to organize your items. The tray itself is heavy, durable, and well-made. The compartments and edges seem to be authentic wood, and the base surface of the tray is a rubber-like surface that keeps things from slipping while still looking very nice.
I like this tray enough to put it on display as a part of my home decor. It’s not only useful for organizing some odds and ends, but is also very beautiful and well made.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 12, 2026
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