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Description
Human Fetuin B 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: 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: 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. Plasma: Collect specimens using EDTA or heparin as anticoagulants and centrifuge them at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. The supernatant can be tested or stored at -20℃ or -80℃, but repeated freezing and thawing should be avoided. 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 enzyme conjugate are added sequentially to microwells pre-coated with fetuin B 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 fetuin B 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 Fetuin B ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Fetuin-B is a protein encoded by the FETUB gene in humans. The protein encoded by this gene is a member of the placenta family and belongs to the cysteine superfamily of cysteine protease inhibitors. Fetuins are involved in a variety of functions, including osteogenesis and bone resorption, regulation of insulin and hepatocyte growth factor receptors, and responses to systemic inflammation. This protein is likely secreted by cells. | |||||||||||||||||||||||||||||||||
| 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, and other biological fluids |
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4.6 ★★★★★
Based on 27 reviews
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Product Reviews
★★★★★ 5
A must read for all believers!
Practical advice on developing the higher self to command the lower self to achieve internal happiness that is not dependent on worldly outcomes. If you are an atheist then you might only benefit from the first couple chapters, some of the later chapters are more theoretical supported by logic based on holy scriptures like the Quran, Hadith, or Psalms. This is a must-read for all believers, especially Muslims! Many blessings to Al-Ghazzali, this book is one of my all-time favorites!
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Reviewed in the United States on February 1, 2018
★★★★★ 5
is a great scholar if Islam who consolidated the tenets of the ...
Imam Gazali, also known as Hujjatul Islam, is a great scholar if Islam who consolidated the tenets of the religion against corruption. The Alchemy of Happiness is a must read for anyone who wishes to take the journey for self explotation, to better understand the self; and by doing so, establish a solid relationship with Allah, the lord of the heavens and the earth and everything in between. The true happiness will only come through such a relationship.
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Reviewed in the United States on March 9, 2016
★★★★★ 3
Weird translation
Format: Paperback
Good content but the translation is a bit off. Dont know if I can trust the information in it completely. Still okay for the price i guess
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Reviewed in the United States on June 19, 2025
★★★★★ 5
A letter of Gratitude to Irv Yalom for Creatures of a Day.
Format: Hardcover
Dear Irv Yalom,
This letter is a declaration of gratitude for your newly published book, Creatures of a Day, and the artful legacy you’ve bestowed upon the field of psychotherapy. In the era of fast technology and mass production, your attention to relationship and the handcrafted nature of therapy is a life line. In all of your 50 years as a psychotherapist, you didn’t sell out for clinical blueprints and formulaic approaches. Instead, you opted to stay true to what you knew--the here and now, the importance of the therapeutic relationship, and your own internal thoughts and experiences as essential elements for your work with clients. Creatures of a Day waves a flag and asks us to take notice. It invites us back into the mystery of our work and reminds us to celebrate our humanness. Your masterful story telling allows us to see you and your clients in action, mistakes and vulnerabilities included, and shares pivotal moments that will provoke thoughtful learning for generations of therapists. So thank you for this.
You show us your mistakes
You see, your books (especially Love’s Executioner and Creatures of a Day) let us into your thoughts and experiences. We get to hear you talk to yourself and occasionally grapple with doubt. We get to know your own vulnerabilities and how they influence your therapeutic relationships. This is such a rare view. And I am hungry for it. There are too few books, too few videos, and fewer workshops or trainings that offer this kind of perspective for therapists. We don’t get to see masters make mistakes. We don’t get to hear supervisors or consultants narrate their doubts. So, I consider your book an invitation to write about my own similar encounters in my work. And to continue to make this kind of conversation central to my trainings and retreats for therapists. Thank you for the inspiration and the permission.
You offer central themes and an individualized perspective
As a collection of psychotherapy tales, I think of Creatures of a Day as a series. Like a series of paintings that are created around central themes, your tales invite us to look at the existential themes of aging, death, and connectedness. And, just like a painting series, each reader will take with them a message that is individualized and of unique importance to him/her. In this way, you are truly exhibiting your art as a writer and a psychotherapist. In Creatures of a Day, two patients read the same book and take from it a very different, but beautifully applicable, message. A nurse perceives the angry words she hissed to her dying patient completely opposite of the way in which they were received by the woman she was treating. And a case that you filed away as a blunder turns out to have been a life changer that is only revealed about a decade or so later. This is a reminder to me that while we can’t predict how our art is received, we can in fact commit to creating and collaborating in the very best way we can.
You invite humanness and the art of relationship
I’d like to let you know that in addition to the invitation to write about my experience as a therapist, I welcome your permission to be human with my clients. And, with that comes a renewed dedication to knowing and experiencing what being human is for me. This means deepening my relationship with my art, continuing my work in therapy, and showing up with the same honesty and openness that you let us see in your book.
You make risk a good thing
You ask your patients to risk and use this in as a very important subject during the course of treatment. You take several risks in Creatures of a Day, and show us that risks are a vital part of being an authentic and real therapist. You show us that in your work you are just being honest and attending to your experience and the client’s experience. In fact it is more risky to be untruthful or hide than it is to show up and attend the the relationship.
You inspire me
I won’t stop practicing. You inspire me to continue to write about my own work. And in my own small way, carry your legacy forward.
Once again, thank you for your guidance, your influence, and your legacy.
Lisa Mitchell, MFT, ATR, LPC
www.innercanvas.com
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Reviewed in the United States on March 31, 2015
★★★★★ 5
Short book, big recommendation!
Format: Hardcover
Great book, smooth read that is not overly technical for those who aren't therapists themselves. Examines large existential questions in a digestible format with each chapter being a different real world story. Highly recommend.
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Reviewed in the United States on April 6, 2026