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Description
Human FS 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 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. 3. 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 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 follistatin (FS) 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 follistatin (FS) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Follistatin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Follistatin (FS), also known as activin-binding protein, is encoded by the FS gene. Follistatin is an autocrine glycoprotein expressed in nearly all tissues of higher animals. It is produced by follicular stellate cells (FS) in the anterior pituitary gland. It is a member of the TGF-β superfamily. FS increases follistatin levels by increasing muscle mass in certain core muscle groups, thereby increasing life expectancy. The relationship between follistatin and polycystic ovary syndrome is also under investigation. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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4.6 ★★★★★
Based on 642 reviews
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Product Reviews
★★★★★ 5
This is really a great read for individuals wanting to increase their knowledge of ...
There was so much information in this book that has never been bought to light. This is really a great read for individuals wanting to increase their knowledge of the Black Power Movement.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 20, 2016
★★★★★ 5
Review of Deacons for Defense
Format: Paperback
I found this book to be extremely informative about the Deacons. I was in Monroe ,LA in 1965 as a staff lawyer for the Lawyers Constitutional Defense Committee. We represented Deacons in various instances. This history filled in innumerable gaps in my knowledge of the Deacons. To the extent of my recollection the facts and analysis seemed accurate to me. This book is recommended for anyone with an interest in acquiring a full understanding of important civil rights organizations which contributed to the Movement in the '60's'
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 3, 2014
★★★★★ 5
I love this book
Format: Paperback
I love this book. Some of the incidents took place in my home town of Jonesboro, LA. I knew many of the men (and women) who risked their lives to take a stand for racial equality. CORE's Jonesboro Freedom House (although it is vacant) still stands today, and its structure has not changed. This house, along with many of the pictures of individuals, are shown between pages 107 & 108 of the book. I shared this information with some of my students. They were quite surprised that our small town was a part of History!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 14, 2017
★★★★★ 5
Funny, Interesting, Informative!
Format: Paperback
I hiked the PCT in 2016 and I wish this had been available then. There are a lot of misconceptions about the desert section of the PCT (first 700ish miles) being flat, monotonous, and unpleasant. Even once you are out on the trail hiking through varied terrain, it is hard to shake off the "desert mentality" that comes from listening to those misconceptions. One of the reasons I love this book is because of how well the author paints an accurate picture of the beauty and constantly changing scenery the desert has to offer.
All the information you would expect from a guide book is here, including mileage, water sources, elevation, etc. There are also a TON of photos that will make you question why you spend anytime indoors. I mean this literally. This book is worth buying for pictures of the Sierra sections alone. The damn near turn by turn navigation descriptions of each section are also a good reason for your purchase.
The author is funny. I wanted to say that in a funny way but she's funny enough so if you want to chuckle, wait for the book. Her passion for the PCT seeps through her writing and it is impossible not to feel equally enthusiastic. On a multi-day (or multi-month) hike, that enthusiasm is almost as important as your next water source...except not..but ALMOST.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 18, 2017
★★★★★ 5
Anyone Can Do It!
Format: Paperback
My husband and I are hiking the PCT and this book has come in very helpful. We especially appreciate the areas we can park at the beginning and end of each section. It is making the trail very doable for two older people who could never make it otherwise. I tear out the section, make a copy and highlight some of the details for example a gate at 4.5 miles. It helps encourage us to continue when we know about how far we have left to go. Also the little map showing elevation gain and lost. The last 17 mile trip we did last week was predominantly up hill. Knowing that ahead of time prepares us mentally for a difficult hike. Our next section is 10 miles downhill so we will do that as a day hike and bring the dog. For anyone who thinks they can't do the PCT this is the perfect book. If I can do it Anyone Can Do It!
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Reviewed in the United States on February 14, 2022
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