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Description
Human 1433PR 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 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 14-3-3 protein (1433PR). After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of 14-3-3 protein (1433PR) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human 14-3-3 protein ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | 14-3-3 proteins are a family of conserved regulatory molecules expressed in all eukaryotic cells. They bind to a diverse array of signaling proteins, including kinases, phosphatases, and transmembrane receptors. Over 200 signaling proteins have been identified as 14-3-3 ligands. Elevated levels of 14-3-3 proteins in cerebrospinal fluid may be a sign of Creutzfeldt-Jakob disease. | |||||||||||||||||||||||||||||||||
| 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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4.3 ★★★★★
Based on 680 reviews
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Product Reviews
★★★★★ 5
Great computer stand
Color: Gray
These stands are great. We now have two of them. My wife suffers from Carple tunnel syndrome and these stands lesson the effects when typing. They are stable and hold the computer securely.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 29, 2025
★★★★★ 4
Weighty and well designed
Color: Gray
The Bestand Laptop Stand is a testament to quality and thoughtful design.
From an ergonomic perspective, this stand truly shines. It elevates my laptop screen to eye level, promoting better typing and viewing to reduce strain on my back and neck.
Another standout feature is the weighty and solid construction of this stand. It's crafted from a single piece of aluminum, which not only adds to the stand sturdiness but also gives it a sleek, modern look. The surface is spacious and accommodates laptops from 11 to 16 inches.
Both the weight of the stand and the silicone treads below and underneath the laptop, and on a metal lip in front, ensure that neither the stand nor the computer move around. The manufacturer claims a 66-pound load-bearing capacity for the stand, and I believe it. This thing is rugged.
The inclusion of a slot at the rear for cables to pass through is a small yet incredibly practical detail. It helps keep my workspace organized and tidy.
Three slots in the stand aid with heat dissipation and are yet another thoughtful design feature that cools my laptop during extended usage.
While the stand excels in many respect, one limitation is its lack of height adjustability. However, if height adjustability is important to you, you’ll find lots of other options for stands here on Amazon.
Bottom line: I like it. Chances are, you will too. The Bestand Laptop Stand is a nice blend of durability, functionality, ergonomic comfort, and attractiveness—all at a reasonable price.
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Reviewed in the United States on September 25, 2023
★★★★★ 5
Pleasantly Surprised
Color: Gray
Honestly, I was a little hesitant buying a laptop stand that didn't offer an adjustable angle, but for the price, I couldn't pass up checking this out. I'm very glad I did.
First, this stand came really well packaged. For a $37 stand, I wasn't expecting much, but box and packaging was impressive. After pulling the stand out of the box, my first impression was this was both weighty and well machined. The edges were smooth, there were no imperfections I could find and the space gray color was dead-on.
As far as functionality goes, this is a tremendous improvement form having my MacBook Air sitting directly on my desk; the angle is perfect for my use case, the wiring hole in the back is more than adequate and the whole setup just looks slick. If you're hesitant about buying a cheaper stand that doesn't offer an adjustable angle, don't worry about this one, it does the trick.
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Reviewed in the United States on October 24, 2021
★★★★★ 5
Upscale look
These valet trays have a really upscale look and feel. I originally thought they’d make a great Father’s Day gift, but after using them, I realized they’re perfect for everyday use by the front door for mail, keys, wallet, and other essentials.
They have a nice weight and perfect size to them and the felt bottom keeps surfaces protected from scratches.
The combination of acacia wood and metal gives them a modern, sophisticated style that works well in an office, bedroom, entryway, or nightstand area. They’re both practical and decorative, making it easy to keep things organized while still looking polished.
Nice quality product.
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Reviewed in the United States on May 12, 2026
★★★★★ 3
Missing screws, can’t contact seller
The trays are really nice, however one came with missing screws. I’ve tried to contact seller via phone number that Amazon provided however the number is disconnected. Great quality other than the missing screws.
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Reviewed in the United States on April 30, 2026
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