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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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4.2 ★★★★★
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★★★★★ 5
Great Product
Size: 180 Count (Pack of 1)
Really appreciate these. Just signed up for monthly delivery
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Reviewed in the United States on March 18, 2026
★★★★★ 5
Great for bad immune systems
Size: 180 Count (Pack of 1)
I used to be sick a MINIMUM of 5x a year. Coming up on a little over a year and a half of consistent use and I have only been sick ONCE
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Reviewed in the United States on March 22, 2026
★★★★★ 5
Best Magnesium source!
Flavor Name: Unflavored, Size: 90 Count (Pack of 1), Flavor Name: Unflavored, Size: 90 Count (Pack of 1)
I have tried many forms of Magnesium in the past, it’s an essential mineral for the body (regulates over 300 functions). It helps with sleep, blood pressure, cognitive function, mood, muscle cramps, and ATP production (the powerhouse of the cell).
Science talk aside, I have tried Magnesium Glycinate and Citrate, and they can do a number on the gut, moving things along too quickly! Magnesium L-threonate has become very popular, but I am very picky about my supplements and the companies that make them. Jarrow Formulas is top notch!
I have started looking for supplements that are more “bio-available” as they are better absorbed and used by the body. This supplement does all of the above, without upsetting my gut and making it overactive.
I ran out of this supplement a few months ago, money was really tight, and it really affected my sleep, which is priceless! It also has L-threonate, which promotes sleep and positive mood. I am happy to have this back in repertoire, combined with vitamin D once a week (50,000IU), and Calcium Hydroxyapatite (see my other reviews, also super bioavailable).
If you are looking for a Magnesium source that helps you sleep and regulate body functions without the side effects on your gut, this is a standout choice that works great!
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Reviewed in the United States on January 4, 2026
★★★★★ 5
My Monthly Magnesium Supplement
Flavor Name: Unflavored, Size: 90 Count (Pack of 1)
I have been ordering the Jarrow Formulas Magnesium L-Threonate, MagMind on repeat every month because it has become an essential part of my routine. I take it consistently to support brain health, memory, and overall cognitive function, and I’ve noticed it helps me feel more focused and mentally sharp. The capsules are easy to take, and the supplement seems to absorb well, which is important for magnesium effectiveness. I appreciate that it’s high-quality and reliable, which is why I keep coming back every month without hesitation.
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Reviewed in the United States on March 7, 2026
★★★★★ 5
Jarrow is a trusty brand, Magmind is superior for cognition, spatial relations and SLEEP QUALITY - UPDATED REVIEW
Flavor Name: Unflavored, Size: 90 Count (Pack of 1)
I have been using Jarrow Formula supplements for a number of years, especially the Vitamin B12 - methylcobalamin and Vitamin K2 - MK7 supplements. I have been impressed with their continuing development of new *effective* products at a great price point.
My husband and I have been taking approximately a gram a day of supplement Magnesium (cheaper products) for almost 5 years now. In that time we have cured his former type 2 diabetes (after almost a 2 decade battle with medicated fasting BGs in the 500s at one point!) and we have almost completely reversed his hypertension of many years as well has have turned his end stage Severe Congestive Heart Failure around so he's relatively free of symptoms. I credit not only the magnesium, but also high doses of Vitamin D (ALWAYS taken with magnesium and fish oil!!), a whole foods source multi, the Sinatra Protocols for supplements as found in the back of
and a number of minerals especially trace minerals as found in
.
I was looking into research on magnesium and the brain when I ran into the MIT study published in 2010 showing magnesium L-threonate is the only form of magnesium to cross the blood-brain barrier and showed observable improvements in cognitive measures. Interesting. So we bought a bottle at our local neighborhood nutrition shop after a discussion with the owner, a couple I have come to trust during our many battles with the medical establishment.
Jarrow Formulas of magnesium L-threonate as Magmind is a very cost-effective source. To further economize we are still taking our regular magnesium supplement 3 times a day (750 mg) then taking this form of magnesium as part of our bed time routine (replacing the 4th magnesium of the day). I am genuinely impressed. Our sleep has improved yet another notch, which is saying something since the 5 years of magnesium supplementation meant we already had really good sleep quality. What I have noticed is that after multiple head trauma injuries (including a serious concussion earlier this year) my previously very high level of spatial relations was slipping. Now with Jarrow I feel I'm getting that edge back. I am very impressed. I will update again with further results after we've been taking this formula longer.
2015 UPDATE - in the years since we first tried MagMind I've referred hundreds of people to try this quality supplement with great results. We've since learned to couple magnesium with taurine for even greater results. Taurine is VERY well documented to improve the fluidity of electrolytes within the cells including within the brain.
NOVEMBER 2016 UPDATE:
Well, here we are 4 years later. We still personally use and recommend MagMind. What we discovered though, since he was put on Armour Thyroid by our primary care he no longer seems to be "leaking" magnesium hence we've cut magnesium supplementation waaayy back. Plus we soak in epsom salts far more. Magnesium is still vital however we're no longer "taking a gram a day."
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Reviewed in the United States on November 15, 2012