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Description
Human CTSB 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. 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. 3. 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. 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 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 B (CTSB) 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 B (CTSB) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Cathepsin B ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Cathepsin B (CTSB), encoded by the CTSB gene, belongs to the lysosomal cysteine protease family and plays a crucial role in intracellular proteolysis. It is produced in muscle tissue during metabolism. It can enhance the activity of other proteases, including matrix metalloproteinases, urokinase (serine protease urokinase plasminogen activator), and cathepsin D. Consequently, it plays a crucial role in proteolysis of extracellular matrix components, disrupting intercellular communication, and reducing the expression of protease inhibitors. It also participates in autophagy and catabolism, which are beneficial in tumor malignancy and may contribute to specific immune resistance. Its overexpression is associated with invasive and metastatic cancers. Elevated CTSB levels in various diseases contribute to numerous pathological processes, including cell death, inflammation, and the production of toxic peptides. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.1 ★★★★★
Based on 10 reviews
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Product Reviews
★★★★★ 4
Small, bu holds a lot
Color: Black
I initially purchased the Jewelry Box Necklace Ring Storage Organizer as a gift for my 8-year-old daughter. The compact design and practical features seemed perfect for her little trinkets. However, its quality and style so took me upon receiving it, I decided to keep it for myself!
The jewelry box is small, but it holds a lot with a double-layer structure that provides ample storage space for various types of jewelry. It easily accommodates rings, necklaces, and other small accessories without clutter.
One of the highlights of this product is its portability. Its compact size makes it an ideal travel companion. I've taken it on several trips, and it has proven incredibly convenient. It keeps all my jewelry neatly organized and easy to find, eliminating the hassle of untangling necklaces or searching for a matching earring.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 1, 2023
★★★★★ 5
Small but nice
Color: green
Cute small jewelry box—
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 21, 2026
★★★★★ 5
Perfect travel mate!
Color: Black, Color: Black
I love this small travel jewelry case. It is a great size for a vacation anywhere. I like how everything has a place to keep you well organized. Great value for the money and good quality material. so easy to snap shut and put in your suitcase. Sleek and black color that is classy.
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Reviewed in the United States on December 12, 2025
★★★★★ 3
Closing it when you have a ring.
Color: Black, Color: Black
Its a nice box but the problem i have with it is the middle piece. It does not give enough room if you have a ring that sits high like a cocktail ring. And same if you have a pendant necklace in the necklace area. You have ro really force it closed and i worry that it will put pressure on the items. It would work better without that middle piece for earrings or maybe if it were half the size. Closes fine when its empty but the minute you put a ring in the ring spot that has any kind of height you have to squeeze it shut. Its a good little box that you can put in a safe. Thats why i use it. Just to put my good jewelry in it and then secure it properly in a small house safe. Also ive already had the backs of a few earring pop off when I've opened the earring flap because it all rubs together. .
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Reviewed in the United States on February 14, 2026
★★★★★ 5
Super usefull
Color: pink
If you're in search of a stylish and functional way to organize your jewelry while on-the-go, then this pink travel jewelry organizer is definitely worth considering. With its compact design and thoughtful features, it's the perfect accessory for any traveler or jewelry lover.
One of the standout features of this organizer is its size. With a compact design that can easily fit into a purse or suitcase, it's perfect for taking your jewelry with you on the go. And despite its small size, it still offers plenty of storage options - from compartments for necklaces and bracelets to slots for rings and earrings, it can hold all of your essential pieces.
But beyond its size, this travel jewelry organizer is also incredibly stylish. The soft pink color adds a feminine touch, while the sleek design and gold-tone hardware give it a sophisticated look. It's the perfect accessory to carry with you on any trip or to use as a daily storage option for your jewelry.
Another great feature of this organizer is its thoughtful design. The compartments and slots are all padded to protect your jewelry from scratches and damage, while the zippered closure ensures that everything stays secure and in place. Plus, the compact design means that you won't have to worry about your jewelry getting tangled or jumbled together during travel.
Overall, if you're in need of a travel jewelry organizer that is both stylish and functional, this pink option is definitely worth considering. With its compact size, ample storage options, and thoughtful design features, it's a great investment that will help you keep your jewelry organized and protected while on the go.
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Reviewed in the United States on April 22, 2023