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Description
Human LTF 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 pre-chilled PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled PBS and resuspend them in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the level is very low, the PBS volume can be reduced 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. Breast milk cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis, or store at -20°C or -80°C, but avoid 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 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 lactoferrin (LTF) 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 lactoferrin (LTF) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Lactoferrin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Lactoferrin, also known as lactoferrin transfer protein (LTF), is a multifunctional protein of the transferrin family. Lactoferrin is a globular glycoprotein with a molecular mass of approximately 80 kDa and is widely present in various secretions, such as milk, saliva, tears, and nasal secretions. Lactoferrin is also present in the secondary granules of PMNs and is secreted by some apical cells. Lactoferrin can be purified from milk or recombinantly produced. Lactoferrin is a component of the human immune system. It possesses antimicrobial (bactericidal and fungicidal) activity and is part of the innate defense, primarily in the mucosa. Lactoferrin interacts with DNA and RNA, polysaccharides, and heparin, and exhibits some of its biological functions in complex with these ligands. | |||||||||||||||||||||||||||||||||
| 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, tissue homogenate, cell lysate, breast milk, cell culture supernatant and other biological fluids |
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4.9 ★★★★★
Based on 1563 reviews
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Product Reviews
★★★★★ 5
I Noticed Improved Digestive Results
I really liked these Migcopat fiber gummies. I started taking them after a gap from my last supplement and noticed a change. I felt a positive impact on my digestive results. My bowel movements were easier to pass, cleaner, and more well-formed. The pectin-based gummies were a little above average size, but they were soft and easy to chew. They did not stick to my teeth at all. They were sugar-free. They came in a clear plastic bottle. The gummies had a tasty, natural blueberry flavor. A serving size was two or three gummies, giving twenty or thirty servings per container. The formula included biotin, keratin, zinc, apple cider vinegar, and other vitamins. I was very happy with the taste, the texture, and the results. The Supplement Facts print was small on the container - it was a little larger on the box.
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Reviewed in the United States on May 23, 2026
★★★★★ 5
Great blueberry aroma, easy to chew, sugar-free formula, long expiration date
The blueberry aroma when I open the jar is wonderful, and the taste is excellent. I really like that the texture breaks down pretty easily when chewed instead of sticking to my teeth. It is great to find a sugar-free option that packs in fiber, prebiotics, and probiotics, plus a few extras like biotin and collagen. I have been taking supplements like this for years, so I do not expect to feel any drastic changes, but everything is working well for me with no ill effects. It is also nice that the expiration date is clearly printed on both the box and the bottle, giving my batch about 23 months of shelf life.
Pros
✅ Excellent blueberry taste and aroma.
✅ Soft texture breaks down easily without sticking to teeth.
✅ Sugar-free formula with a great 23-month expiration window.
✅ Includes hair and skin support ingredients like biotin and collagen.
Neutrals
– The 60-count bottle yields 20 to 30 servings depending on whether you take two or three gummies.
Cons
❌ None observed, these are very pleasant to take.
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Reviewed in the United States on May 12, 2026
★★★★★ 5
Tasty, Easy to Take
This is the 3rd flavor of this brand that I've tried. I have to say that I preferred the grape and the little strawberry hearts better than these, as far as taste. These also are quite high in Biotin at 33,333% of the daily value, if you take the 3 gummies that equal 10g fiber. If you do want 10g of fiber the jar won't last an entire month. I don't really need the hair/nail support, so I'll probably go back to one of the other flavors. These are so easy to get out of the wide-mouth jar and they don't stick together too much. They're a soft gummy and are easy to chew. I don't tend to have an issue with them sticking to my teeth. The jar I received was manufactured in Apr 2026 and expires Apr 2028. Another reviewer mentioned that they're made in the USA, but I don't see that claim anywhere in the listing or on the product...although they're distributed by a New York company.
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Reviewed in the United States on May 13, 2026
★★★★★ 4
Fewer doses per jar, but good flavor
I've been using migcopat fiber gummies for a while now, and I'm a fan of the brand, so I thought I would give these a shot.
I appreciate the addition of the biotin and keratin.
This just only has one flavor instead of three, which is blueberry, but it's quite pleasant, not overly sweet. These gummies are a little softer than the other migcopat gummies, but again nothing wrong with that.
I am knocking one star from the review because this jerk contains 20 doses when you take three per day as opposed to the other maker gummies that I have where there's a 30-day supply with taking three per day.
For me, the addition of the biotin and keratin doesn't justify losing a third of my supplement.
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Reviewed in the United States on May 21, 2026
★★★★★ 5
Great value
This is 1 of those items that would take months to give a fair assessment. It takes time for things like this to get into your system. This was a vine item and I wanted to try them as fiber is very important after 50. But I can not speak to the effectiveness. I hope and expect there would be no side effects. Texture seems normal for something like this. Not sure if theres a odor. But seems a good value. Not sure about taste either.
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Reviewed in the United States on June 1, 2026