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Description
Human SYT1 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, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a synaptotagmin-1 (SYT1) 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 synaptotagmin-1 (SYT1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Synaptotagmin-1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Synaptobrevin 1 (SYT1) is a protein encoded by the Syt1 gene. Synaptobrevin is an integral membrane protein of synaptic vesicles and is believed to act as a sensor for calcium ions (Ca2+) during vesicle trafficking and egress. Calcium ions bind to synapsin I and participate in triggering neurotransmitter release at synapses. SYT1 is the master switch responsible for neurotransmitter release in the brain. It can sense calcium ion concentrations as low as 10 ppm and subsequently signal the SNARE complex to open the fusion pore. It has been shown to interact with SNAP-25, STX1A, and S100A13. | |||||||||||||||||||||||||||||||||
| 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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4.6 ★★★★★
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Product Reviews
★★★★★ 5
Great pillow and company
Color: Crescent White, Size: King
I have a couple of Coop Home pillows and loved them. They are several years old and I was on the search for a replacement. I tried 2 other brands, one cost more and the other cost less. Returned 1 and used the other for a short time before I had to go back to my old Coop pillow. I started looking at crescent pillows and did some research. This type of pillow is great for side and back sleepers. Being a side sleeper I looked into the best rated. Coop Home crescent pillow was not rated #1 but it was close. The top rated pillow cost 2.5 times more and I could not justify paying that. I am happy to report that I am waking up less (before every 1.5-2 hours). It comes with extra fill and I love a very firm pillow so I emailed customer service to see if I could get more filling. This was on the weekend and in 1 day they responded that they would be happy to. Now that is great customer service. I sleep warm and have no problem with a hot head. It comes with a cover but I use silk pillowcases and it works fine. I will say when I first got the pillow it was very thin and flimsy. I had some doubts but followed directions and put in the dryer on low. Came out perfect. Highly recommend this pillow and company.
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Reviewed in the United States on April 20, 2026
★★★★★ 5
Best pillow I've ever owned
Color: Cut-out White, Size: Queen
While the pillow looks anemic when it first comes out of its packaging, it puffs/firms up beautifully after time in the dryer. I haven't had any odor issues.
I'm mostly a side sleeper and at first I thought there wasn't enough of the extra fill, so I put it all in. I've ended up needing to take at least half back out to keep my head at the correct angle.
While this can fit in a standard pillow case, it just barely fits and keeps coming out - at least at my fill level. Queen pillow cases are much better.
This is so much more comfortable than any of my previous pillows. I don't need to stack multiple pillows to get the right support. I'm no longer waking up with neck or shoulder or arm pain. I'm recommending it to pretty much everyone and I've purchased a second pillow to keep at my parents for when I visit!
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Reviewed in the United States on May 6, 2026
★★★★★ 5
So nice your wife will steal it for herself
Color: Original White, Size: Queen
This, this is the god of all pillows I’ve ever purchased. It’s the perfect balance of firm and soft and it’s easily adjustable using the added filler. I’m a side sleeper so having a primary pillow is a must. Combined with a decent secondary pillow this one provides a perfect amount of support. This pillow is so nice, my wife ‘borrowed it’.
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Reviewed in the United States on May 27, 2026
★★★★★ 5
Favorite pillow!!
Color: Original White, Size: Queen
Absolutely love this pillow! Bought my husband one a year ago after he was waking up with neck pain. Just recently bought myself one, we both love them! Very comfy and comes with a bag of extra foam to add/remove from the pillow. The zipper on the side of the pillow cover is hidden but durable. We take it with us wherever we go, and always sleep so good.
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Reviewed in the United States on May 30, 2026
★★★★★ 5
These pillows are way nicer than hotel pillows!
Size: Queen / Standard, Color: White
This is a real review and I am not a robot. I’m actually sleeping with these pillows right now. They are so comfortable the right softness levels. Oh my gosh they’re awesome. I don’t know about hotel pillows, but these are way better quality form with your head and you sleep like a baby.
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Reviewed in the United States on May 29, 2026