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Description
Human IRF5 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 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 Interferon Regulatory Factor 5 (IRF5) 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 Interferon Regulatory Factor 5 (IRF5) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Interferon Regulatory Factor 5 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interferon regulatory factor 5 (IRF5) is a protein encoded by the IRF5 gene. It is a member of the interferon regulatory factor (IRF) family, a group of transcription factors with diverse roles, including viral-mediated activation of interferons and regulation of cell growth, differentiation, apoptosis, and immune system activity. IRF family members are characterized by a conserved N-terminal DNA binding domain containing tryptophan (W) repeats. Alternative splice variants exist that encode different isoforms. The regulatory and inhibitory regions of the IRF family are primarily located at the C-terminus of the IRFs. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.2 ★★★★★
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Product Reviews
★★★★★ 4
Works great for dry skin!
I stumbled across this product when looking for a new lotion for my 2 month olds super dry skin. I tried several lotions until finally this one worked. It is fragrance free which is a huge plus. It was vent gentle on my babies skin and helped with her dry skin fairly quick. This is a staple in our nighttime routine now! Only thing I do not love about it, is that it is a little thin/liquidly which I personally don't like. Overall, great product and would re buy!
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Reviewed in the United States on May 11, 2026
★★★★★ 5
Great Baby Lotion!
Love this lotion for my baby. It keeps her skin soft and smooth and absorbs quickly. I even use on myself at times as well.
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Reviewed in the United States on February 23, 2026
★★★★★ 5
Amazing for sensitive babies
This lotion is amazing!! My baby girl has extremely sensitive skin nothing worked for her and other well known brands kept creating irritation for her. Her pediatrician recommended this brand and it truly soothes her skin with no irritation, moistures beautifully all day long and has a light refreshing scent! It’s a must have for babies that are extra sensitive! Pairs well with the body wash as well.
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Reviewed in the United States on April 24, 2026
★★★★★ 5
The best foundation!
Style: Refill, Color: Golden Glow, Style: Refill, Color: Golden Glow
I recently tried the Jane Iredale makeup powder foundation and I have to say, it's absolutely amazing! This foundation has quickly become my go-to for achieving a flawless and natural-looking complexion.
One of the things I love about this powder foundation is its lightweight. It feels very light on the skin and allows it to breathe, which is perfect for those who prefer a more natural and comfortable makeup look. It doesn't feel heavy or cakey at all, even after hours of wear.
The coverage of this foundation is also impressive. It provides just the right amount of coverage to even out my skin tone and conceal any imperfections, while still allowing my skin to look like skin. It doesn't settle into fine lines or emphasize dry patches, giving me a smooth and radiant finish.
Another great feature of this foundation is its long-lasting power. It stays put throughout the day, even in humid weather or during physical activities. I don't have to worry about it melting off or fading, which is a huge plus for me.
I use the Jane D2O spray prior to putting on the powder and a final spritz after applying to set.
I also appreciate the fact that Jane Iredale powder foundation is made with high-quality and skin-friendly ingredients. It is free from harsh chemicals and irritants, making it suitable for sensitive skin types. It gives me peace of mind knowing that I'm not compromising the health of my skin while wearing it.
Overall, I feel Jane Iredale foundation is a game-changer. I highly recommend it to anyone!
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Reviewed in the United States on March 7, 2024
★★★★★ 5
The best foundation!!!
Style: Refill, Color: Golden Glow
10/10 powder foundation. If you’re on the fence, just do it. I have used this product since 2014 and haven’t ever bothered to try and find anything better. There’s simply no need. I’ve used the color Golden Glow almost every time, and it has always managed to match my skin throughout all the seasons. It is lightweight but full coverage. It’s magic. I mostly wear this on its own, with just moisturizer as a base. It’s so easy to apply with a large brush, and run out the door feeling like you have a full face of makeup on. It does pair beautifully with my liquid foundation for when I’m looking for maximum coverage. The price point is fair in my opinion. Each refill last me about 6 months. You do have to purchase the compact case which is an addition $25 but that’s a one time fee. I have never been happier or purchased a skincare product more frequently than this one. You seriously won’t regret it.
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Reviewed in the United States on January 13, 2026
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