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Description
Human TRAM1 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 ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected 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 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 capture antibody against Translocation Associated Membrane Protein 1 (TRAM1). 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 Translocation Associated Membrane Protein 1 (TRAM1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Translocation Associated Membrane Protein 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Translocation-associated membrane protein 1, also known as TRAM1, is encoded by the TRAM1 gene. It participates in the translocation of nascent protein chains into and through the endoplasmic reticulum (ER) membrane by promoting correct chain positioning within the SEC61 channel. It regulates the exposure of nascent secretory protein chains to the cytosol during translocation into the ER. It may affect the phospholipid bilayer near the lateral gate of the SEC61 channel, thereby promoting ER protein trafficking. Throughout the integration process into the ER membrane, it closely associates with the transmembrane (TM) domain of nascent membrane proteins. It associates with the second TM domain of nascent chains of the G protein-coupled receptor opsin/OPSD in the ER membrane, potentially facilitating membrane integration. Under ER stress conditions, it participates in the disposal of misfolded ER membrane proteins during the unfolded protein response (UPR), an integrated stress response (ISR) pathway, by selectively translocating them from the ER to the cytoplasm where they are located, leading to proteasome ubiquitination and degradation. | |||||||||||||||||||||||||||||||||
| 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 and other biological fluids |
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4.3 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 5
The topics deals with so many areas that the body of christ may face.
Format: Paperback
I love this book by Pastor Brenda. Once you read this book you can apply it to so many areas in you life.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 6, 2024
★★★★★ 5
God's Will for Every Situation
Format: Paperback
We all want God's Will! but do we always look for it...try to find it ...search for it... pray for it ... and for every situation?
Well, Brenda Kunneman has really addressed all these questions and so many more in this important-to -read book she has written.
Following the Introduction this great author leads us directly to Chapter One" 'You Can Know God's Will', and then of importance she teaches in Chapter Eleven 'Finding God's Will For Finding A Church'. This is on prime importance. I know of one church where the main pastors are carrying for years now un-forgiveness in their hearts and are still preaching and teaching. Wow!
This is just a great book for all to read and it is a great book for small group studies along with the Bible. Read and grow in the Lord.
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Reviewed in the United States on October 1, 2011
★★★★★ 5
Great fit and makes potty training easier
Size: 3T-4T, Unit Count: 84
These training pants have been a big help during potty training. They fit comfortably and feel soft against the skin, more like real underwear than a diaper. The stretchy sides make them easy for little ones to pull up and down on their own, which really helps with independence.
They do a good job holding small accidents without leaking, and the fun character designs make my little one more excited to wear them. The pack comes with plenty to last through the training phase, which is nice when you’re going through a few each day.
Tip: keep a few extras in the car or diaper bag for outings. It makes potty training on the go so much easier.
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Reviewed in the United States on October 14, 2025
★★★★★ 5
Great Pull-ups that Don't Leak!
Size: 3T-4T, Unit Count: 20
I was on the hunt for the perfect potty training pants for my little guy, and Pull-Ups Boys' Potty Training Pants in size 3T-4T turned out to be a great find. We’ve tried a few different brands, but these have been the most reliable. The design is fun and colorful, which my son loves, and it makes him excited to wear them. The fit is spot on for his size (32-40 lbs), and he moves around comfortably without any leaks. The absorbency is impressive – it gives me peace of mind knowing accidents won’t mean a complete outfit change. Overall, these Pull-Ups have made the potty training process a lot smoother for both of us.
There are a few things to note, though. The price is a bit higher compared to some other brands, but I believe the quality justifies it. Also, while they are great during the day, we still need a bit more protection at night. I find myself using an extra layer for nighttime just to be safe. The packaging could also be improved – a resealable option would be nice to keep the remaining training pants fresh and organized. Despite these minor issues, I’m really happy with the product, and my son is well on his way to being fully potty trained.
Pros:
- Fun and colorful design that my son loves
- Perfect fit for 32-40 lbs, allowing comfortable movement
- Excellent absorbency, minimizing leaks
- Helps make the potty training process smoother
Cons:
- Slightly higher price compared to other brands
- Not as effective for overnight use, requiring additional protection
- Packaging lacks a resealable option for convenience
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Reviewed in the United States on June 5, 2024
★★★★★ 5
Leak proof training pants
Size: 3T-4T, Unit Count: 112
These training pants are amazing. My son uses them whenever he’s taking a nap or sleeping throughout the night. I’ve ordered these multiple times and we’ve never had an issue with leaking.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 19, 2026
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