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Description
Human aFP 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. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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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| Sensitivity | 0.071 ng/mL | |||||||||||||||||||||||||||||||||
| 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 an alpha-fetoprotein (aFP) capture antibody. 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 alpha-fetoprotein (aFP) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Alpha-Fetoprotein ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Alpha-fetoprotein (AFP, also sometimes called alpha-fetoprotein, alpha-1-fetoprotein, alpha-fetoglobulin, or alpha-fetoprotein) is a protein encoded by the AFP gene. The AFP gene is located on the q arm of chromosome 4 (4q25). Maternal AFP serum levels can be used to screen for Down syndrome, neural tube defects, and other chromosomal abnormalities. AFP is the major plasma protein produced by the yolk sac and fetal liver during fetal development. It is considered the fetal analog of serum albumin. AFP binds to copper, nickel, fatty acids, and bilirubin and exists as monomers, dimers, and trimers. | |||||||||||||||||||||||||||||||||
| 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 homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.2 ★★★★★
Based on 16 reviews
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Product Reviews
★★★★★ 5
A good sun solution for sensitive lips!
Color: White, Size: 0.15 Ounce (Pack of 1)
Having sensitive lips can be super frustrating. They burn SO easily in the sun. But most sunscreen lip balms on the market irritate them. Those that don't irritate aren't moisturizing at all, so my lips become dry and chapped. Which leads to more sensitivity. Ugh. But so far this Sun Bum SPF 30 has worked for me! No sunburnt lips. No irritation. Enough moisture to keep my lips soft and healthy while working or playing outside. Sure, it's s not really pleasant tasting when it gets in your mouth, and like most zinc-based sunscreens, it makes my lips milky white. But it works for my picky lips and it's a good value for the money--it's become my summertime staple.
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Reviewed in the United States on July 20, 2023
★★★★★ 3
Hydrates, leaves a slight white film, not travel friendly
Color: White, Size: 0.15 Ounce (Pack of 1)
I wear lip balm regularly. As a matter of fact, I always have some with me. Tubes are placed in various areas around my house, I have one in my car, in my backpack and a couple tubes at work. I'm seriously never without it so I kind of consider myself a lip balm expert. The only thing most of the ones I use don't have is SPF. Since I live in an area that is fairly sunny year round, you think I'd be better about that. Well, I'm looking to change that.
I saw this Sun Bum SPF 30 Mineral Sunscreen Lip Balm and read some of the reviews. They seemed overwhelmingly positive. I'm familiar with the brand and since it was covered under my HRA, decided to try this one out along with a few others. It glides on easily and moisturizes well, but it left a slight white film on my lips. Not my favorite but it did fade. It's unscented and doesn't have any flavor, which is good. I loathe lip balms with any type of taste to them. Good or bad. It has clean ingredients and offers great sun protection but it doesn't withstand heat itself. If you like to carry one in your pocket, backpack or keep in your car, it will melt.
I do like this lip balm but it has some negatives, as stated above. It made my lips feel soft and hydrated but it's not travel friendly. I wouldn't discourage buying it, but if you want a balm to carry around with you, I'd be hesitant to recommend it.
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Reviewed in the United States on February 26, 2024
★★★★★ 5
Really works and it's safe.
Color: White, Size: 0.15 Ounce (Pack of 1)
I am a flats fisherman – this stuff really works, it's smooth, flavorless, reef-safe, and safe to lick. Reapply as often as you remember to.
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Reviewed in the United States on March 22, 2026
★★★★★ 5
Highly recommend. Any graininess and whiteness are minimal and insignificant.
Color: Blanco, Size: 0.15 Ounce (Pack of 2)
This is the best mineral(non-chemical) lip sunscreen I have found. Highly recommend. The first few times I used it, it felt ever so slightly grainy. Maybe the graininess rose to the top. Even if it was grainy all the way through, I would still recommend. There is a slight whitish cast to your lips-not noticeable or significant in my opinion. There is a very minimal flavor and it has a nice feel on your lips.
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Reviewed in the United States on February 7, 2026
★★★★★ 5
A must have for your summer bag
I am 100% devoted to Blue Lizard products! I got some extra sun and needed relief, fast. This non-greasy nor slimy cream is exactly what I didn’t know I was looking for. I had applied aloe alone to my skin and received no relief. Within minutes if not seconds of applying this, I had noticed improvement. The consistency is not too thick or too thin, it absorbs into the skin nicely and can be reapplied without getting cakey. It gave my sunburn relief while also moisturizing the skin. I haven’t peeled at ALL. I love that it has clean ingredients and a neutral fragrance. My skin is so sensitive, so having natural ingredients is a plus. Blue Lizard After Sun is now my holy grail and I will recommend it over and over.
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Reviewed in the United States on June 8, 2026