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Description
Human PHB 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 and centrifuge 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. Cell Supernatant: 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 and mince the tissue. 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 the 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. Preparation before testing: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the gradient working solution of the standard: 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. 3. 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 as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 4. 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. 5. 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. 6. 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 a prohibitin (PHB) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the prohibitin (PHB) 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 Prohibitin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Prohibitin, also known as PHB, is a protein encoded by the PHB gene. PHB genes have also been described in animals, fungi, plants, and unicellular eukaryotes. Prohibitins are divided into two classes, designated type I and type II, based on their similarity to yeast PHB1 and PHB2. Every organism possesses at least one copy of a single type of prohibitin gene. Prohibitins are evolutionarily conserved and ubiquitously expressed. The prohibitin gene, located on the BRCA1 chromosome at 17q21, was initially thought to be a negative regulator of cell proliferation and a tumor suppressor. This antiproliferative activity was later attributed to the 3' UTR of the PHB gene, rather than the actual protein. Mutations in PHB have been associated with sporadic breast cancer. However, overexpression of PHB is associated with decreased androgen receptor activity and reduced PSA gene expression, leading to reduced growth of androgen-dependent prostate cells. Levels of the longer transcript are higher in proliferating tissues and cells, suggesting that this longer 3' untranslated region may function as a trans-acting regulatory RNA. | |||||||||||||||||||||||||||||||||
| 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, cell supernatant, tissue homogenate, etc. |
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4.7 ★★★★★
Based on 19 reviews
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★★★★★ 1
Flimsy Nozzle
Size: 2 fl oz
The nozzle is so flimsy at the moment I pressed on it it popped off the can into the gas tank. I didn’t press it softly but I also didn’t press it very hard either, getting it out was a huge pain in the butt.
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Reviewed in the United States on May 24, 2026
★★★★★ 5
The Ultimate Multi-Use Motor Treatment – A Must-Have for Any Vehicle!
Color: Pack of 4
The Sea Foam SF-16 Motor Treatment is an outstanding product that truly delivers on its promises. After using it in multiple applications for my vehicles, I’m convinced this is a must-have for anyone looking to maintain and enhance engine performance. The 4-pack is an excellent value and ensures I always have some on hand for routine maintenance.
Why Sea Foam Stands Out
Multi-Use Versatility: This product is a game-changer! I’ve used it as a fuel additive, an oil treatment, and even to clean out stubborn deposits in my carburetor. It works flawlessly across applications, making it an all-in-one solution for keeping engines running smoothly.
Improved Performance: After adding Sea Foam to my fuel tank, I noticed a significant improvement in my vehicle’s fuel efficiency and a smoother idle. It’s particularly effective at removing deposits and buildup that can reduce performance over time.
Easy to Use: The instructions are straightforward, whether you’re using it for fuel, oil, or cleaning purposes. The precise measuring cap ensures no mess or guesswork, making it simple to get the correct dosage every time.
Great for Long-Term Maintenance: Regular use of Sea Foam has made a noticeable difference in prolonging the life of my engine. I’ve also used it to revive an older lawnmower engine, which now runs like new! This product pays for itself by preventing costly repairs.
Packaging and Value
The 4-pack is perfect for multi-vehicle households or anyone maintaining more than just a car. I’ve used it in my truck, car, and small engines like my generator and lawn equipment, and I still have plenty left.
Pro Tips
Add Sea Foam to your fuel before a long drive to maximize its cleaning effect.
Use it regularly in oil changes to keep sludge buildup at bay.
It’s also great for seasonal storage—just add it to your gas tank to stabilize fuel and protect the engine.
Final Thoughts
Sea Foam SF-16 is the real deal for anyone who values vehicle performance and longevity. Its multi-use functionality, ease of application, and proven results make it a standout product. I’ve already seen noticeable improvements in my vehicles and equipment, and I’ll continue to use it as a staple in my maintenance routine. Highly recommend it to anyone who takes pride in keeping their engines in top shape!
Pro Tip: Whether you’re a DIY mechanic or just looking for a reliable maintenance product, Sea Foam is a must-have. It’s one of the simplest and most effective ways to protect your investment and extend the life of your engine. Don’t hesitate—this 4-pack is worth every penny!
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Reviewed in the United States on January 12, 2025
★★★★★ 5
Engines run smoothly and efficiently
Color: Pack of 2
I’ve been using Sea Foam SF-16 in both my outboard motor and sailboat, and it has proven to be an invaluable product for maintaining engine performance. This motor treatment is incredibly versatile and effective, making it a staple in my marine maintenance routine.
One of the standout benefits of Sea Foam SF-16 is its ability to clean and lubricate the entire fuel system. It effectively removes harmful deposits and residues that can build up over time, ensuring that my engines run smoothly and efficiently. This is particularly important for marine engines, which can be prone to issues due to the harsh operating environment.
I also appreciate how Sea Foam stabilizes fuel. This is crucial for my sailboat, which sometimes sits unused for extended periods. By adding Sea Foam to the fuel tank, I can be confident that the fuel remains fresh and free from moisture, preventing potential engine problems when I’m ready to hit the water again.
Another great feature is its ease of use. Simply adding the recommended amount to the fuel tank is all it takes to start seeing the benefits. I’ve noticed a significant improvement in engine performance, with smoother idling and better throttle response. It also helps reduce emissions, which is a nice bonus for the environment.
Overall, Sea Foam SF-16 is a fantastic product that I highly recommend to anyone with an outboard motor or sailboat. It keeps engines clean, running smoothly, and protected from the elements, making it a must-have for any boat owner.
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Reviewed in the United States on September 24, 2024
★★★★★ 5
WOW-Smoother Performance-Idle Better-Easy To Use-LOVE This Stuff
Color: Pack of 2
GREAT product! I own a "mature" car with lower milage. However the low idle was off and slow, additionally learned that a Gas station provider (occasionally use) had been found to have water in their gas. After reading the reviews and learning many auto mechanics swear by Sea Foam, decided to give it a try. As a woman have experienced hype, un-necessary work etc through the years. I used a funnel to add Sea Foam to my gas tank. Wow! even on shorter trip, noticed a smoother performance. With longer driving seems to have helped the idle and running even smoother! Believe that with more use will see even better performance and hopefully better milage! Do Not Hesitate, you and your vehicle are worth it! Thank you!
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Reviewed in the United States on December 19, 2025
★★★★★ 5
Good stuff for small engines
Color: 3 Pack
I use this for my lawn equipment and outdoor tools. Keeps them running great.
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Reviewed in the United States on May 12, 2026
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