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Description
Human LTbR 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 Lymphotoxin Beta Receptor (LTbR) capture antibody. 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 Lymphotoxin Beta Receptor (LTbR) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Lymphotoxin Beta Receptor ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Lymphotoxin beta receptor (LTbR), also known as tumor necrosis factor receptor superfamily member 3 (TNFRSF3), is a cell surface receptor for lymphotoxins involved in apoptosis and cytokine release. The protein encoded by this gene is a member of the tumor necrosis factor (TNF) receptor family. It is expressed on the surface of most cell types, including epithelial and myeloid cells, but not on T and B lymphocytes. LTbR not only helps trigger apoptosis but also leads to the release of the cytokine interleukin-8. Overexpression of LTbR in HEK293 cells increases IL-8 promoter activity and leads to IL-8 release. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.3 ★★★★★
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Product Reviews
★★★★★ 5
The pads have changed but still a good product
I have been using these make up remover pads for many years now and will continue to use them. I like the 3 pack so that I do not run out. I usually only use 1 pad a day so a 3pk will last me for a long while. I do not have any issues with the pads drying out in the time frame from opening them to using the last one so that is a major plus. I have no issues with these on my skin or that close to my eyes and they do a great job of removing my mascara, which is the only make up I use. I recommend them. I will say that in 2025 they have changed the material for their pads, they are now thinner and do not have the texture they used to have which for me the only difference is that they can be a bit more difficult to separate but nothing to complain about.
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Reviewed in the United States on November 7, 2025
★★★★★ 5
Excellent Product!
Ordered for my Lady, she’s says it’s an Excellent Product fir Removing Eye Make-Up.
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Reviewed in the United States on May 2, 2026
★★★★★ 5
Amazing make-up remover pads, that work just as well as oil base ones, without the oil.
I have tried many many make-up removers and I always come back to this one. I stumbled onto this one at a pharmacy a long time ago, and I can honestly say it's the best. I have tried many others since, high end, lower end, but this is my favorite. it's obviously extremely convenient, being in a pad, rather than having to wet a pad yourself, which poses a host of problems. Sometimes you run out of pads and you have to use toilet paper, which sucks, sometimes it spills, which you them have to clean, making an annoying two step process even more annoying, by it being a three step process. These convenient pads, a one step process, are so great, because unlike most other pads I tried, these are moist. I tried so many pads that lose their moisture after you open the package, and these remain very moist. I love that its a twist off cap. Also, they are the best at removing eye makeup, better than anything I tried, except for oil pads, HOWEVER, unlike oil pads, these do not make your vision blurry and get into your eye. This brand also makes oil pads, which i dislike. Oil always manages to get into my eyes, but the thing about oil pads is that they work at removing makeup. Well Guess what?!?! These work as well, WITHOUT THE OIL!!! And they DON'T dry out your eyes and face. Love them.
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Reviewed in the United States on September 13, 2015
★★★★★ 1
Ouch!! Update: Mold inside?
Update: I reduced my three star review to one star due to finding black specs all over lid inside my second jar with some black specks on the top pads. Mold? Mildew? I don't know have mold in my cabinets and I keep this in my top vanity drawer which is on its own wall nowhere near the tub or shower. I was so disgusted I threw out the whole jar plus my third one. I hate to write a bad review on a product that is such a good value and so needed in this market. Maybe it's just a one-off but either way I will not be repurchasing due to the soapy, stinging pads - mold or no mold. My conscience is telling me to warn people.
Original review:
They sting like there is micellar water in them? Plus soap. They are very sudsy and make you feel like you have to rinse your eyes before they dry out, which defeats the purpose. They do work well to get mascara off but so does soap. Sadly will not replace the old Almay oil-free remover pads which I have used for years. Have not found a good replacement for my dry, sensitive eyes. I will have to use them on other parts of my face but not my eyes.
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Reviewed in the United States on January 10, 2024
★★★★★ 4
My last batch has changed - be aware!
I have used these pads for several years now. They are generally very good and do the trick. I don't even look at other products because they are so good.
However, in this last order, they no longer have a smooth side and a bumpy side. They always had the different texture which helped in using them. I agree they are a little small which I have gotten used to, but now with both sides being smooth, they are much harder to use to wipe. Plus you don't have the advantage of the texture. This is a dowgrade for sure.
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Reviewed in the United States on October 22, 2025