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Description
Human TRAF5 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. 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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| Theory | This kit utilizes a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP enzyme conjugate are sequentially added to microwells pre-coated with a TNF Receptor Associated Factor 5 (TRAF5) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of TNF Receptor Associated Factor 5 (TRAF5) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human TNF Receptor Associated Factor 5 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | TNF receptor-associated factor 5 (TRAF5), also known as RING finger protein 84 (RNF84), is a protein encoded by the TRAF5 gene. This gene encodes a scaffold protein that is a member of the tumor necrosis factor receptor-associated factor (TRAF) protein family and contains an alphaprotein and TRAF homology (MATH) domain, a RING-type zinc finger, and two TRAF-type zinc fingers. TRAF proteins associate with members of the TNF receptor superfamily and mediate their signaling. This protein is a component of a multiprotein complex that binds to the cytoplasmic domain of the tumor necrosis factor (TNF) receptor and mediates TNF-induced activation. | |||||||||||||||||||||||||||||||||
| 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 homogenates and other biological fluids |
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4.2 ★★★★★
Based on 26 reviews
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Product Reviews
★★★★★ 4
It Works
Size: 1 Fl Oz (Pack of 1)
I think this Product works and rates high on yuka.👍
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 2, 2026
★★★★★ 5
make my dark spot lighter quickly
Size: 2 Fl Oz (Pack of 1)
I decided to order this product only after reading numerous reviews. I’ve already used it twice, and I feel that the dark spots on my face have faded significantly. Perhaps due to the demands of raising a child, my complexion had become quite dull; however, this product is incredibly gentle—it leaves my skin feeling neither greasy nor dry, and it doesn't have a strong scent. I think I will be repurchasing this indefinitely. I can't wait to see my skin continue to improve!
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Reviewed in the United States on April 5, 2026
★★★★★ 5
Makes skins smooth.
Size: 1 Fl Oz (Pack of 1)
Great product for skin.
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Reviewed in the United States on May 22, 2026
★★★★★ 3
BEST? (the "R" is for relative)
I have, to date, tried four different Retinol serums (usually containing hyaluronic acid). Each of these has been in a similar price range and none of them qualifies as high-end pricing. From what I can tell each of them is as effective as the other and none of them produces "miracle elixir" results. Like most maintenance products they are only truly effective through consistent long-term use. Seeing that I have found each of the brands I have tried to be equally effective I believe it is fair to say that this brand of Retinol serum is acceptable. However, this 1 oz. bottle is one of the higher priced brands I have tried and the bottle was almost half empty when I received it. It is also perfumed and thicker than other versions and therefore harder to measure effectively (one drop is almost sufficient but two drops is too much). When you reach the bottom of the bottle, like with most of the other brands I have tried, it is impossible to get the last quarter inch of product out of the bottle using the dropper. The syringe is not long enough or flexible enough. What I have been doing is either adding a few drops of water or a few drops of Vitamin C serum to liquefy the balance so it can be shaken out onto your finger. Although use of water will obviously dilute the serum it at least allows you to use all of the product, which is a compromise I am willing to make. As recommended by most Retinol serum brands, and vice versa, I have been applying Vitamin C serum after using it. However I am not convinced that use of both products is really necessary, i.e., Retinol serum combined with a Vitamin C serum. The only sources I see strongly promoting the advantage of using both are the sources that are selling both.
As someone commented in response to my initial post (which I have modified since I wish to be accurate), Retinol is not hyaluronic acid. However many Retinol products contain hyaluronic acid which is what caused my confusion.
"Skin care products with hyaluronic acid are most frequently used to treat wrinkled skin although they don't replace anything the body has naturally lost. These are very effective moisturizers." Retinol is made from vitamin A, and it's in many non-prescription skin care products. Retinol's stronger counterpart is tretinoin, which is the active ingredient in Retin-A and Renova, available by prescription only. If your skin is too sensitive to use Retin-A, Retinol is an alternative, although the effects are less impressive. Retinol may improve mottled pigmentation, fine lines and wrinkles, skin texture, and skin tone and color." (Both quotes excerpted from WebMD.)
"Retinol is photosensitive and should only be worn at night. Never go outside or into the sunlight with it on; this could cause adverse effects." (Huffington Post quote) (Using a sunscreen is strongly recommended by many sources because retinoid products cause increased photosensitivity.)
Having researched this a bit more I feel certain that I can stop using Retinol serums altogether since my dermatologist prescribes a stronger product for evening use. I will ask him about this the next time I see him, though. He already advocates use of a Vitamin C serum but has never commented on the benefits of using it in combination with a Retinol product. However, since Retinol is photosensitive I believe eliminating it would be okay and probably to my benefit since sunscreens wear off after a while and need to be reapplied. If he advises on this subject in any way I believe might be helpful to others I will add it to my post.
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Reviewed in the United States on June 22, 2016
★★★★★ 5
Approves
Size: 2 Fl Oz (Pack of 1)
I’ve been using it for about two weeks I’ve noticed an improvement in my wrinkles and skin texture
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Reviewed in the United States on May 7, 2026