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Description
Human SASH1 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 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, biotinylated detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a capture antibody against SAM and SH3 Domain Containing Protein 1 (SASH1). 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 SAM and SH3 Domain Containing Protein 1 (SASH1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human SAM And SH3 Domain Containing Protein 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | SAM and SH3 domain-containing protein 1, also known as SASH1, is a protein encoded by the SASH1 gene. This gene encodes a scaffolding protein involved in the TLR4 signaling pathway, which may stimulate cytokine production and endothelial cell migration in response to invading pathogens. The encoded protein has also been described as a potential tumor suppressor that may negatively regulate cancer cell proliferation, apoptosis, and invasion, and reduced expression of this gene has been observed in various cancers. Mutations in this gene may be associated with abnormal skin pigmentation in patients. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.8 ★★★★★
Based on 536 reviews
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Product Reviews
★★★★★ 3
Assembly is harder than it should be; you get what you paid for
Color: Black, Size: 4 Panel-88''
Review of SUNALLY Room Divider 6FT Folding Privacy Screens, 4 Panel Fabric Divider for Room Separation, 88" W Freestanding Portable Wall Dividers Screen for Home Dorm Studio Office, Black
I hate when I have to leave products a not so good review...
Assembly is harder than it should be, mainly because there shouldn't BE any assembly. I have ordered several room divider privacy screens over the last couple of years and not one of them needed any kind of assembly. With that aside, it STILL was a pain in the sphincter to assemble. One of the "legs" just sits on top of a foot plate - it's not screwed in to anything.
It's kind of flimsy and the base plates don't have anything on them to not scuff up the floor.
With all that said, though, it does stand up and will block out what I need to block out. At the less-than-$50 price-point, it's kind of what should be expected. If I had purchased this outright, I would have considered sending it back and getting a higher quality. But if you only have $50 to spare and don't mind assembling things, it could be good for you.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 28, 2026
★★★★★ 5
Great buy
Color: Black, Size: 4 Panel-88''
This room divider is exactly what I needed. When you have 2 kids sharing a room and don’t want to spend a lot of money to divide their space this is what you can use. Believe it or not they are much happier now. This came in a small box which scared me but once I was able to get this up I was very pleased. It was easy to put together. The fabric is pretty good quality, it’s stable and stands up fine. Good value and worth it to have peace and quiet in the house.
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Reviewed in the United States on May 18, 2026
★★★★★ 1
Junk
Color: Black, Size: 4 Panel-88'', Color: Black, Size: 4 Panel-88''
Horrible. The parts are not right. It won’t fit together or disassemble and can’t return it like this. Parts won’t fit properly and missing holes for screws. Also, can’t get through to a person for any assistance. Picture shows the hole and then the lack of a hole where a screw belongs.
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Reviewed in the United States on May 13, 2026
★★★★★ 5
Good product, poor directions.
Color: Black, Size: B-88''W-4 Panel
The quality of the product was better than most at this price. The directions could have been much better even though I have an engineering background.
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Reviewed in the United States on September 1, 2025
★★★★★ 4
Nice partition for my basement
Color: Beige, Size: B-102''-3 Panel
This 3-panel room divider showed up well-packaged with all the parts and even a little tool for putting it together. The instructions were easy to follow, and while one person can definitely assemble it, having two makes it quicker and less frustrating.
It does exactly what I needed—gives quick, easy coverage in an open area. I'm using it in my basement to hide the furnace and water heater, and it works great for that. It looks nice overall, but I picked the beige fabric (which is really more of a tan), and the black frame makes it stand out more than I’d like. A white or lighter frame would’ve blended better. Not sure that I would use it anywhere besides the basement
It’s lightweight and pretty sturdy. The wheels make it easy to move, but it does get a bit wobbly, and the panel clips need to be readjusted every time you roll it somewhere new. Still, for the price, it’s a solid buy. I’m happy with it and planning to order another one.
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Reviewed in the United States on June 21, 2025