Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Jul 8 - Jul 13
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
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. |
|||||||||||||||||||||||||||||||||
| 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 |
|
|||||||||||||||||||||||||||||||||
| 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. |
|||||||||||||||||||||||||||||||||
| 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 |
Shipping Notes
- Free Standard Shipping on $100+ Orders to the USA.
- Except Preorder products are shipped in 48 hours.
- Delivery to the USA:
- Standard Shipping : 3-10 business days
- If time is of the essence, please consider selecting expedited delivery for faster service.
Exchange/Return Notes
- We offer a 30-day return/exchange service after receiving.
- Final sale items are not eligible for returns or exchanges.
- To process your return/exchange, please contact us at [email protected]
- Please click here for more details>>> Return & Exchange Policy
4.6 ★★★★★
Based on 1006 reviews
Sort
Product Reviews
★★★★★ 4
Excellent book, but NO chapters between books in the index
Format: Hardcover, Format: Hardcover
Excellent book, with a complete introduction that covers different points of view of the creation of such work of art, it can be seen that it was translated with a team of high academic preparation and knowledge on the subject. It was translated with a lot of precision, respect, professionalism and with much seriousness to the subject. The book is made with high quality and its pages are quite thick. The only mistake that I could notice and it is difficult for me not to be upset, is that they created the index and left out the chapters of each book. For me it is a gross error in comparison with other versions. It's the reason why I could not give 5 stars.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 9, 2019
★★★★★ 5
A much needed translation that turned out beautifully
Format: Hardcover
Two things make this translation of the Picatrix a must have for any fan of the work. First, it has an index, and it's a really nice index that is fairly complete. Second, the paragraphs are numbered, so we can refer to things as being mentioned in Picatrix 3.4.1, and then somebody else can go look it up and say, "Ha ha, good one." The translation seems to be really solid, too. It's a great edition.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 11, 2019
★★★★★ 5
Burnout meet your enemy
Format: Paperback, Format: Paperback
Soul Care offers a refreshing perspective on managing burnout and finding balance in today’s fast-paced world. Through its insightful guidance, the book explores how to savor life's small moments, connect deeply with oneself and others, and cultivate a meaningful relationship with God.
Debra masterfully addresses the concept of burnout, providing practical advice on how to pace oneself and integrate rest into daily routines. By emphasizing the importance of taking small, manageable steps, the book empowers readers to make gradual but impactful changes in their lives.
One of the most valuable aspects of Soul Care is its focus on both the journey and the destination. It encourages readers to appreciate the process of personal growth and spiritual development, rather than just striving for an end goal. This balanced approach helps foster a deeper connection with oneself and with God.
The book is rich in actionable insights and offers easy-to-implement strategies that are perfect for those seeking to improve their well-being and spiritual life. Its gentle yet profound advice serves as a beacon for anyone feeling overwhelmed and seeking a path to renewal.
Overall, Soul Care is a compassionate and practical guide that is likely to resonate with anyone looking to rejuvenate their spirit and enhance their connection with God, all while navigating the demands of modern life
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 8, 2024
★★★★★ 5
Discover the Soul-Centered Rhythms of True Self-Care
Format: Paperback
In today's fast-paced world, it can feel nearly impossible to escape the cycles of burnout, stress, and exhaustion. Soul Care by Debra Fileta is a timely and much-needed guide for anyone longing to break free from functioning on empty and embrace a life of true, lasting fullness. As a licensed counselor, Fileta expertly blends biblical wisdom with practical insights from clinical psychology, offering readers a holistic approach to self-care that touches the mind, body, and spirit. What I found especially impactful is how Fileta draws from the life of Jesus to demonstrate six key life-giving practices that are not only restorative but also sustainable in the long term. Her ability to bridge scripture and actionable self-care methods makes this book both spiritually enriching and immediately practical. From debunking myths about self-care to addressing how neglecting your soul affects your relationships, work, and ministry, Soul Care is filled with clear, relevant strategies. Fileta’s guidance on how to proactively address burnout is invaluable, especially for those of us in helping professions or ministry roles where we often pour into others while running on empty ourselves. This book is a breath of fresh air, a reminder that self-care is not selfish but essential for serving others well. If you're looking for a biblical, well-rounded approach to personal renewal, this book is a must-read!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 15, 2024
★★★★★ 5
Great book
Format: Paperback
I really enjoyed this book and author.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 11, 2026