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Description
Rat MUC1 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 and centrifuge at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be collected for analysis or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. Saliva: Tissue homogenate: 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 and mince the tissue. 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 collected for analysis. 4. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 mucin 1 (MUC1) 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 mucin 1 (MUC1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Mucin 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Mucin 1 (MUC1), cell surface-associated, also known as polymorphic epithelial mucin (PEM) or epithelial membrane antigen (EMA), is a mucin encoded by the MUC1 gene. It is a glycoprotein with extensive O-linked glycosylation in its extracellular domain. A member of the mucin family, mucins line the apical surfaces of epithelial cells in the lungs, stomach, intestines, eyes, and several other organs. Mucins protect the body from infection by binding to oligosaccharides in their extracellular domain, preventing them from reaching the cell surface. Mucins are cleaved in the endoplasmic reticulum into two fragments: a cytoplasmic tail comprising a transmembrane domain and an extracellular domain. Its overexpression is often associated with colon, breast, ovarian, lung, and pancreatic cancers. This protein has protective functions by binding to pathogens and also possesses cell signaling capabilities. In the nucleus, it regulates the activity of transcription factor complexes that have a documented role in tumor-induced changes in host immunity. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, saliva, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.6 ★★★★★
Based on 7 reviews
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Product Reviews
★★★★★ 4
Does this beat Anthony Invigorating wash in the long last smell
Scent: Seagrass & Driftwood, Size: 32 Fl Oz (Pack of 1)
Anthony Invigorating wash (check out my review of that) has a very strong pine scent to it and is a very long lasting smell. How does Cremo's products stand up to it!
I put deodorants, shampoos, and body washes through the same rigorous tests. I run 14+ miles 4-5 times a week, swim 20+ lengths (or laps) in a junior Olympic pool 3 days a week, use a Sauna 2-3 times in fifteen minute intervals 4 times a week, and use weights 4 days a week. All of these body washes and shampoos that I try out not only go up against Anthony's but also getting rid of that chlorine smell. I always will finish a product before reviewing it
I'll be honest here as always, the Blue Cedar and Cypress at first whiff smelled weird. I can't explain the 1st whiff smell though you have my warning. I thought this was going to be my least favorite of the 3 different Cremo products (bourbon and oak, seagrass and driftwood, and this one) that I received. I have the policy to use all the products up in their entirety, so you don't have to. After using it up, I actually like the smell of it. The cedar smell comes out once applied. I actually use this with Anthony Blue Sea Kelp Body (check out my review of that). This goes great with it.
So on my buy, borrow, or bury review rating system, this is a buy. The issue I have with it, is that Cremo products don't seem to be that moisturizing their smells are good and lasting. This beats the off the chlorine smell of the pool and works great in combination with Anthony's. Try it out, just get past that initial smell, you won't be disappointed.
Now for Cremo's seagrass and driftwood. The First smell test of it is very pleasant. It's once you've put it on does that wood smell really come out. The smell is good and does last long though the issue I have that it is not moisturizing. This goes great with Anthony's other 2 products that I have viewed (please check those out). So on my buy, borrow, or bury rating scale, this is a buy. If they could make their products more moisturizing it would be fantastic.
I'll be back here to review the Bourbon and Oak, I have not finished it yet though it's probably going to be my favorite of all the 3 Cremo products I've tried out.
Please check out all of my other reviews!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 20, 2020
★★★★★ 5
Great buy!
Scent: Palo Santo, Size: 16 Fl Oz (Pack of 2)
This is my favorite brand and the lather is second to none! The smell is amazing and the quality is worth the extra price. It leaves my skin feeling moisturized and soft.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 4, 2026
★★★★★ 5
FINALLY. THIS IS ACTUALLY WHAT YOU WANT.
Scent: Seagrass & Driftwood, Size: 32 Fl Oz (Pack of 1)
I’ve tried all of the soaps. All of them. Molten brown sucks, 18.21 sucks, Blackwood sucks, and brickell sucks.
All I wanted is a premium soap that doesn’t suck. All I wanted is to feel good and smell good. I work in kitchens, so I’m not some desk monkey needing to wash off the stale air, I have to actually clean myself.
I don’t even know how these soaps stay in business, you might as well be using Irish spring and save the money. Nothing moisturizes, I’m always left with dry skin, or the smell lasts all of 6 minutes.
Until now.
This stuff is actually the luxury brand I’m swearing by. It lasts on my skin, it lathers well, it cleans well, and it doesn’t give me itchy skin just to smell nice after a shower.
Also, it doesn’t cost $36 per bottle like molten brown does for 2 teaspoons of soap with gold flakes in it that smells great for 6 minutes. It’s cheaper, and you get more.
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Reviewed in the United States on September 14, 2024
★★★★★ 5
Long Lasting Scent
Scent: Bourbon Oak, Size: 16 Fl Oz (Pack of 2)
I've tried dozens of different brands of body washes and have had various objections.
This is the best by far in every metric.
First of all the scent. Bourbon & Oak.
I'm not convinced that it smells like bourbon OR oak. It's definitely a warm musky scent and I would say it tends more towards a creamy spice with a strong low note.
Longevity:
My job is about 50 percent physical throughout the day and I tend to shower only at night when I get home from work. This is the only body wash whose aroma not only lasts through the rest of the evening, but well into late morning the next day. I would even say that physical activity brings out a very smooth and subtle character. This is the first time I've ever been able to smell a body wash the next day. It was surprising and pleasant.
As for moisturizing, my skin feels smooth but without the oily residue that I typically associate with moisturizing products.
If a product lives up to my expectations I tend to leave a simple review and fair rating.
For products like this that exceed my expectations I try to leave a more detailed review that highlights its strengths.
I will most certainly continue to use this brand and experiment with the different scents available.
10/10 would recommend.
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Reviewed in the United States on October 29, 2023
★★★★★ 3
Please do NOT reship items like body soap that have been returned
Scent: Seagrass & Driftwood, Size: 32 Fl Oz (Pack of 1), Scent: Seagrass & Driftwood, Size: 32 Fl Oz (Pack of 1)
So I've purchased this twice now, I love the scent and I love how it lathers up, overall very happy with the produce so long as I get a product that wasn't previously returned. The 2nd order I received, it was clear that someone had replaced the soap in the bottle with some other kind of liquid and returned it and I got it. First off that has to be dangerous and hazardous.
As you can see from the pictures, something is wrong with this product.
The first product I got had a clean scent, was clearly a body soap liquid. The 2nd product I ordered was more water, smelled had virtually no scent at all. Unfortunately I had already poured it into my soap dispenser so it's now not returnable. PLEASE DO NOT SHIP OUT RETURNED ITEMS!! It's dangerous
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Reviewed in the United States on February 15, 2024