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Description
Human ASP 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. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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 acetylcholine stimulating protein (ASP) 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 acetylcholine stimulating protein (ASP) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Acylation Stimulating Protein ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Acylation-stimulating protein (ASP) is an adipokine isolated based on its function as a potent fat storage factor. It has three disulfide bonds formed by cysteine residues. It stimulates triglyceride synthesis and storage in adipocytes by enhancing glucose and fatty acid uptake, an action that is independent of and additive to the lipogenic effects of insulin. Knockout of ASP results in increased food intake and energy expenditure. | |||||||||||||||||||||||||||||||||
| 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 homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.7 ★★★★★
Based on 29 reviews
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Product Reviews
★★★★★ 3
Difficult to grasp
Format: Hardcover
I love the aspects of career counseling. But this book makes the theories hard to understand.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 20, 2018
★★★★★ 5
Good
Format: Hardcover
Was great quality
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 8, 2021
★★★★★ 5
High energy mindset book, but very repetitive
Format: Hardcover
The 10X Rule is a very high energy, motivational book focused on taking massive action and setting goals much higher than what most people think is realistic.
The strongest part is the mindset shift. It constantly pushes the idea that success requires way more effort, persistence, and volume than people typically expect. If you are looking for motivation to work harder and avoid playing it safe, the message definitely hits.
It also does a good job challenging excuses and comfort zones. The tone is very direct and intense, which can be useful if you respond well to that style of motivation.
However, the book is very repetitive. The same core idea of “10X effort” is repeated throughout almost every section in slightly different ways, which can make it feel stretched out.
It is more motivational than practical, so if you are looking for step-by-step systems or structured strategies, this may not be what you are expecting.
Overall, it’s a strong mindset book if you want intensity and motivation, but less useful if you are looking for detailed execution plans.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 13, 2026
★★★★★ 5
An Excellent, Eye-Opening Read That Truly Makes You Reflect
Format: Kindle
The 10X rule is an excellent book—engaging, interactive, and surprisingly eye-opening. It doesn’t just present ideas; it invites you to actively reflect on your own behavior, decisions, and patterns. As you move through the chapters, you’re constantly prompted to pause, think, and reassess the way you act and respond in everyday situations.
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Reviewed in the United States on February 8, 2026
★★★★★ 5
Grant Doesn't Hold Back!
Format: Hardcover
What It's All About...
So, the 10 X Rule is a very motivation book about rethinking the way you think about goal setting and the effort it takes to accomplish your goals.
Without giving away the obvious, Grant shares that when we go for a goal like, "I want $100 in the next 30 days to pay my bills." We often fall short of that goal and need to restate it as: "I want $1,000 in the next 30 days to pay my bills." Now, if you do far short (happens often), you will still hit somewhere greater than $100. Therefore, we must always 10 X our goals to hit the goals we desire.
Now, when it comes to effort, because we often underestimate the action it takes to meet our goals... Our new $1,000 goal requires 10 X effort to hit it. So, if you think "To hit $1,000, I need to make 1 sales call each day." you need to 10 X that number and make 10 sales calls to hit your new $1,000 goal. Therefore, we must always 10 X our effort and change our actions to meet out goals.
He also takes times at the end of the book to outline 32 characteristics of successful people that also help in accomplishing these 10 X Goals. Some of those include: Have a "Can Do" attitude, Seek to Solve Problems, Focus on Now, Be Highly Ethical, etc.
At the end of each chapter he asks a set of questions that helps to reinforce the material you just read.
My Take...
Overall, this really juiced me up and got me to rethink the way in which I look at goals and the actions I need to meet them.
When reading such a book, I often get so inspired that I shoot off goals without really appreciating the amount of effort it will take to hit them. I walked away from Grant's book with a firm idea on the big goals I want to hit and a healthier understanding of what it's going to take to get there.
The overall feel of the book was also inspiring because Grant is the type of person that believes in going for goals, even if you don't know exactly how you are going to accomplish them... Just get started. I resonate with that line of thinking.
If you like listening to audio books, Grant does his own reading of the book, which gives a greater sense of emphasis and passion behind the content.
Who should read this...
Entrepreneurs, Business owners, High School and College Students
WARNING: There is some cursing.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 20, 2016