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Description
Rat TSG-6 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. 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 capture antibody against tumor necrosis factor-inducible gene 6 protein (TSG-6). 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 tumor necrosis factor-inducible gene 6 protein (TSG-6) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Tumor necrosis factor-inducible gene 6 proteinELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Tumor necrosis factor-induced protein 6, encoded by the TSG-6 (tumor necrosis factor, alpha-induced protein 6) gene. TSG-6 is a 30 kDa secreted protein that contains a hyaluronan-binding LINK domain a and is therefore a member of the hyaluronan-binding protein family, also known as hyaluronan adhesion proteins. The hyaluronan-binding domain is known to be involved in extracellular matrix stabilization and cell migration. The protein has been shown to form a stable covalent complex with alpha-interactin (IαI), thereby enhancing the serine protease inhibitory activity of IαI, which is important in the protease network involved in inflammation. The expression of this gene can be induced by several signaling molecules, mainly tumor necrosis factor alpha (TNF-α) and interleukin-1 (IL-1). TSG-6 expression can also be induced by mechanical stimulation of vascular smooth muscle cells and has been found to be associated with the synthesis and aggregation of proteoglycans. | |||||||||||||||||||||||||||||||||
| 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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4.6 ★★★★★
Based on 2425 reviews
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Product Reviews
★★★★★ 5
These fit very nicely, are comfortable, and work for my wide feet!
Size: 12 Wide, Color: Brown
Once I learned while shoe shopping in my youth that I needed wide-sized shoes, I wasn't able to easily choose from any shoes I saw in the store. I usually have to go find the wide widths and have to deal with the limited styles they have. Shopping online has been hit and miss, too. Sometimes, shoes are advertised as wide width, but end up still being too narrow.
These shoes fit me just right. They were not too tight at the toes, gave good width at the arch, and the opening was large enough that I didn't have to undo every single lace just to get my foot in. They also look nice, and I'm sure will take a decent polish.
Also, note that this does say it's an Oxford shoe, but it does have the open lacing and is more of a derby-style shoe. This also helps with sizing for my wide feet. If I didn't already have a pair of black shoes, I'd look at picking one of these up.
My only reservation with these is that the soles are not dark-colored to complement a more formal style. I feel the white makes them a little more casual. The sole is also more transparent/opaque coloring at the heel and underneath, which you can kind of see-through, so keep that in mind. It's not fully clear, but is kind of transparent, which I was not expecting. I think it also shows where it picks up dirt as you wear it. Just more of a style cue one way or another.
As a reference, I buy the 4E wide and Extra Wides in New Balance and Skechers. I also have a hard time getting any good fit in Nike shoes.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 2, 2023
★★★★★ 5
Looks and feel good
Size: 10.5, Color: Black
Shoes look dressy but feel comfortable. They are stiff but very comfortable.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 8, 2026
★★★★★ 5
Well made and affordable
Size: 10.5, Color: Brown
Very comfortable, great price!!!!!!
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Reviewed in the United States on April 30, 2026
★★★★★ 5
Perfect Blend of Comfort and Style
Size: 9, Color: Brown, Size: 9, Color: Brown
I recently purchased size 9 of these shoes, and they exceeded my expectations in every way! Here’s what I love about them:
1. Comfort: The moment I put them on, I noticed how soft and supportive they felt. The cushioning provides all-day comfort, making them perfect for long walks, workdays, or running errands.
2. Fit: The sizing is spot-on! I ordered my usual size, and they fit perfectly with no need to size up or down. There’s enough room for my toes to move without feeling too loose or tight.
3. Style: These shoes look even better in person. The design is sleek and versatile, making them easy to pair with both casual and semi-formal outfits. I’ve received several compliments already!
4. Durability: I put it on the day of purchase. After wearing them daily for a couple of days, they still look brand new. The stitching and material seem high-quality and built to last.
5. Value for Money: For the price, these shoes deliver exceptional value. You’re getting premium comfort and style without breaking the bank.
Final Thoughts: If you’re looking for shoes that offer comfort, style, and durability, I highly recommend giving these a try. I’ll definitely be considering another pair in a different color soon!
To conclude, I’m very happy for this purchase. Will definitely buy again.
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Reviewed in the United States on January 13, 2025
★★★★★ 5
Great Purchase
Size: 12, Color: Black
These shoes are great. They look great and extremely comfortable.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 10, 2026
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