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Description
Mouse Cath-C ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 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 a Cathepsin C (Cath-C) 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 Cathepsin C (Cath-C) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Cathepsin C ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Cathepsin C (CTSC), also known as dipeptidyl peptidase I, is a lysosomal cysteine exoprotease belonging to the peptidase C1 subfamily of cysteine cathepsins. It is encoded by the CTSC gene. Cathepsin C plays a key role in activating granular serine peptidases in inflammatory cells, such as elastase and leukoplastic enzymes in neutrophils, and chymotrypsin and tryptophanase in mast cells. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.4 ★★★★★
Based on 8 reviews
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Product Reviews
★★★★★ 5
It Does a Great Job!
Style: 51oz｜Straight Styles
This brewer is great for cold brewing loose teas. Is glass and Stainless Steel, so no micro trash.
The glass is very delicate so handling needs to be careful.
Great product. I bought another one.
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Reviewed in the United States on April 16, 2026
★★★★★ 5
Makes Fantastic Cold Brew. Thoughtful Design. Slightly Frail, But Approaches Perfection.
Style: 51oz｜Conical Styles
When I looked for a dedicated cold brew pitcher, I had a few conditions. Mainly I wanted to avoid plastic at all costs. I needed a pitcher that would be easy to clean and simple to use, without pointless bells and whistles. Finally, I wanted a pitcher that could fit in my fridge without issue.
This pitcher achieves all that and then some. Borosilicate is the same stuff Pyrex used to make its glassware from, a material renowned for its durability especially when exposed to quick temperature shifts. I knew it was overkill, but such material would surely fit the bill. And glass wouldn't affect the flavor of whatever coffee I'd brew in it.
The grind holder is made from stainless steel, with a fine mesh that largely prevents sediment from getting into the coffee. It does allow smaller particles to get through, but they settle to the bottom. Overall I can't complain.
The grind holder is suspended by a rubber ring which also locks the pitcher's lid in place. This forms a nice little seal that seems to help prevent leaking. Obviously there is no vacuum capability here, but it feels sturdy and like it won't let anything in or out once the grind holder is in.
Pouring is easy and accurate. I remove the lid and grind holder, then pour. There's never any spillover nor dripping. The pitcher has a good, well-crafted pouring tip.
Brewing with the pitcher takes a bit of practice. First I dump old grinds if necessary, then fill the grind holder roughly 3/4 of the way with cold brew grinds. BEFORE I place the grind holder into the pitcher, I fill the pitcher with water about 2 inches below the max line. After that, I place the grind filter into the pitcher then slowly press down on the grinds to flatten and wet them. Finally, I slowly pour water in to hit the max line, flattening the grounds as I do.
While the pitcher has notches to determine how much liquid it holds, I don't find that feature helpful because I only use the pitcher with a grind holder which boosts the liquid. I imagine the notch lines may be more helpful for someone using this as a standard pitcher without the grind holder in place.
I've accidentally overfilled this pitcher many times before deciding the best way, in my opinion, to handle it as described above. I've tried placing the grind holder in and then pouring all of the water over the grinds. But because grinds have a tendency to rise and overflow, that method proved messy and ineffective. This is why I add most of the water first, then only add a very small amount of water AFTER putting the grind holder into position just to top it off.
So how good is the cold brew? After trying a few different approaches, I now brew mine in the fridge for convenience, letting it steep for about a week to make a nice dark roast concentrate with Bizzy Organic Dark & Bold grounds. The result is stellar, easily better than any coffee place cold brew I've had in the past couple of years.
Filled to the max line, a full pitcher lasts me about 2 weeks when I have about an inch of coffee per day. I've found cold brew concentrate to be an extremely economical way to stretch costly grinds while creating the richest flavor and this pitcher is a key part of that. With this approach, I'm a happy customer.
One thing I like to do while brewing a new batch of coffee is pouring about 1/3 of the brewed coffee into a tall cup and finishing that while brewing. This method has worked perfectly for me, as I have a new pot of coffee by the time I'm done with the old one.
One thing to point out is that the pitcher is fairly big and the glass is relatively thin. And so I have always hand washed all parts of this pitcher, which has proven straightforward enough. While borasilicate and stainless steel should be dishwasher safe, this has become too important a piece of equipment for me to start playing around with. So I'm not the person to say how well it holds up in a dishwasher. My outlook is to treat this pitcher with the care it deserves and to be rewarded with its offerings for as long as possible.
Overall the pitcher is an all-star at making the coffee I enjoy, being so simple to use that it becomes easy to take for granted. It is almost a trouble-free piece of kitchen hardware and has already paid for itself through sheer convenience. The quality definitely feels top notch, although the thinness of the glass means it needs to be treated with the care and respect it deserves.
I would feel very comfortable recommending this product to almost anyone serious about making a quality cold brew at home.
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Reviewed in the United States on October 29, 2020
★★★★★ 4
Better than expected (Updated)!
This is a simple and effective solution to making a good cold brew. Too bad the process takes forever (no fault to the product).
BUT if you want a stronger taste, after using purified cold water (try to get beans wet), let it sit out for 12 hours or longer but then place in the fridge and you'll notice it darker and richer.
It is impossible if you think some ground beans won't end up in your coffee, they will BUT when you pour, as long as you leave the metal ring in, a little bit of coffee (water) and the beans usually get stuck and don't come out. If you're really nuts about it, you can use a cheese cloth but a bit of grounds won't hurt you and not sure you'd really taste it as it's so fine, barely noticeable.
Those who keep braking the glass? You must use cold water if you immediately start cleaning the pot. Do NOT run under hot water while the pot is cold, you or course will shatter the glass at some point.
Update (07/29/2020)
I am still using this product and no issues with it. But it does leave the coffee grounds smell to it no matter what beans I use, so wash with soap after every use.
Be sure to know, water will evaporate and also get soaked up in the coffee beans, so adding a little more water above Max may be helpful. Also, pro tip, when you take out the insert with the beans, put it over the lid and tilt it and let it rest for 3 Mins so all the water from the beans pours out, do NOT force it with your fingers as you'll get more grounds and will have a harsh flavor.
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Reviewed in the United States on May 12, 2020
★★★★★ 5
Great for iced tea and easy to use
Style: 51oz｜Conical Styles
I use this for making iced tea and it works perfectly. You can use tea bags or loose-leaf tea, and both come out great. It’s easy to fill, strain, and clean, and I like that I can just keep it in the fridge and pour as I go. The airtight seal keeps everything fresh. Super convenient and a staple for my fridge now
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Reviewed in the United States on November 3, 2025
★★★★★ 5
Great for tea!
Style: 51oz｜Straight Styles
Love this pitcher! Use it for my NORA pregnancy tea to steep overnight in the fridge.
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Reviewed in the United States on May 2, 2026
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