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Description
Human IGFBP-2 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 and centrifuge 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. Urine: Tissue homogenate: 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., 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 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 an Insulin-Like Growth Factor Binding Protein 2 (IGFBP-2) 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 Insulin-Like Growth Factor Binding Protein 2 (IGFBP-2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Insulin Like Growth Factor Binding Protein 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Insulin-like growth factor binding protein 2 is a protein encoded by the IGFBP2 gene. The protein encoded by this gene can be secreted into the blood, where it binds IGF-I and IGF-II with high affinity, or it can remain intracellularly, interacting with many different ligands. High expression levels of this protein promote the growth of several tumor types and may predict a patient's chance of recovery. Several transcript variants of this gene have been identified, one encoding a secreted isoform and the others encoding a non-secreted isoform. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, urine, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.3 ★★★★★
Based on 1315 reviews
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Product Reviews
★★★★★ 5
Beef Tallow Sunscreen
That dry, stretched sensation that I sometimes get from using other sunscreens was not something that I experienced with this one. On the contrary, I had the impression that my skin remained supple and pleasant throughout the morning. I used it prior to going about my errands, having a drive, going for a stroll outdoors, and spending time at the park. To my satisfaction, I did not feel the need to quickly apply another moisturizing lotion on top of the one I was using in order to restore the natural feel of my skin. Simply said, it made it simpler for me to utilize on a daily basis. When I wore it for a longer period of time, I also paid attention to how it felt on my face. Even though I wore it on a number of days when the temperature was high, I did not experience any discomfort as the day progressed. Despite the fact that I was aware that I was wearing sunscreen, I did not feel any discomfort or want to wash my face straight away with it. Because I am quite particular about the products that I use on my face, I found that to be really helpful. In addition to having a sunscreen that is dependable, I want it to be something that I can really endure for a considerable amount of time.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 9, 2026
★★★★★ 5
The Best Way to Keep My Skin Happy Under the Sun
I am so happy that I found this special white cream because it is the best helper for my face and arms when I am playing outside in the big bright sun. I keep the bottle right near my sink so I can put it on before I go out to play, and it is very easy to use because the cream feels very smooth and soft like butter when I rub it on my skin. Sometimes other white creams feel very chalky or yucky like flour, but this one is a great deal for the price because it blends in so well and it does not leave a big white ghost face on me once I rub it in. It feels very simple and clean on my skin and it does not have a scary or strong smell, which makes me very happy because it does not feel greasy or oily like a piece of pizza. Even when I put more on later in the day, it does not clog up my skin or feel heavy like a thick blanket, so it is a wonderful value because I can use it all day long and stay comfortable. The tube is a perfect size to throw in my backpack or my travel bag when we go on a trip, and it feels very strong and tough like it will last a long time without the cap breaking off. It is a very practical and smart tool for my skin because it forms a strong layer that I can see at first to make sure I am safe, and it feels much better and more dependable than thin lotions that disappear too fast. I think everyone who likes to be outdoors should get this because it is durable, it works perfectly to keep me safe, and it is so simple and easy to use that it never gets in the way of my fun!
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Reviewed in the United States on April 14, 2026
★★★★★ 5
Comfortable to Wear and Easy to Use Every Day
The texture was the first thing I felt. First application feels rather rich but unexpectedly distributes easily throughout the skin. I was worried it would feel heavy, but it blended in more readily than I thought and didn’t leave my skin feeling too oily afterwards.
I love that you can use it on the body and face too. One product for the entire family is easier and saves room when packing for trips or holidays. I have used it on myself and my kids when we go out and it has been comfy for all of us to wear.
Another thing I appreciate is my skin feels hydrated after using it. Love things that make my mornings easier on hectic mornings. The mixture is smooth and pleasant to wear all day long and does not feel sticky.
The container is functional and simple to throw in a beach bag or handbag. I have brought it to soccer games, trips to the playground, family picnics and it has been no problem.
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Reviewed in the United States on June 4, 2026
★★★★★ 5
Less cast and the zinc scent is mild.
I use sunscreen regularly. One of the aspects I do not like is the white cast I get from zinc-based SPFs. This one is zinc-based but it has tallow and PDRN. I want to see if they make any difference when it comes application and feel.
MY EXPERIENCE WITH THE PRODUCT
1. I received Mystic Orchard’s beef tallow sunscreen with PDRN. Comparing its value, I think the volume is pretty good. I received 2.8 ounces.
2. When it comes to scent, I smell the metallic scent but it is not as bad as the one I have been using for swimming. It is there, but pretty faint.
3. You can tell by the sheen that it is zinc-based. It has the same consistency.
4. What I noticed though is that the white cast is not too bad. It blends fast with the tallow.
5. I like tallows because they contain fat-soluble vitamins like vitamin E for some antioxidants.
6. The cica is nice too for those who need a calming ingredient with their sunscreen.
7. I think this would be good for when going out. I do not think I will be using it for swimming since it is mild in scent and cast. Better to use it elsewhere than just for swimming.
OVERALL IMPRESSION: I think that this SPF sunscreen with tallow and PDRN is cost-effective and worth its value if you want a zinc-based sunscreen with less cast and milder scent.
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Reviewed in the United States on April 21, 2026
★★★★★ 5
Moisturizing Sunscreen
Size: 2.82 Fl Oz (Pack of 1)
The grass-fed tallow base makes it deeply moisturizing—it feels like a nourishing balm. The PDRN complex is a nice bonus for repair.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 23, 2026
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