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Description
Human TFAM ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 freeze-thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes and remove the supernatant for analysis. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, but avoid repeated freeze-thaw cycles. Pre-Assay Preparation: 1. Remove the reagent 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 stand for 15 minutes to completely dissolve, then gently mix (concentration 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 seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 20ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube is used 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 a Transcription Factor A, Mitochondrial (TFAM) 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 Transcription Factor A, Mitochondrial (TFAM) 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 Transcription Factor A, Mitochondrial ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Mitochondrial transcription factor A (TFAM) is a protein encoded by the TFAM gene. This gene encodes a mitochondrial transcription factor that is a key activator of mitochondrial transcription and a participant in mitochondrial genome replication. It binds to mitochondrial promoter DNA to facilitate mitochondrial genome transcription. It is a double-box, high-mobility group DNA binding and bending protein. This bending function is important for mitochondrial transcription initiation in mammals, but not in its yeast homolog, Abf2. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenate, cell culture supernatant |
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Product Reviews
★★★★★ 5
Great Summer Bridge Activities for Students going into 4th Grade
Good for 3rd going into 4th grade summer activities.
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Reviewed in the United States on April 29, 2026
★★★★★ 5
Perfect for Homeschooling!! Let the stories come alive and captivate you!
Format: Hardcover
I’m a homeschooling mama and I’m grateful for men like Eric Metaxas who are personally invested in the wellbeing of our nation. Thank you, Eric, for sharing history with truth and the fear of God.
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Reviewed in the United States on June 2, 2026
★★★★★ 5
A Revolution of Faith and Freedom
Format: Hardcover
Eric Metaxas’s Revolution: The Birth of the Greatest Nation in the History of the World presents the American founding as far more than a political dispute over taxes or power—it is portrayed as an epic struggle between liberty and tyranny, virtue and corruption, good and evil. Through vivid storytelling and rich historical detail, Metaxas casts the Revolutionary era as a moral drama in which ordinary men were called to extraordinary courage against overwhelming odds in their fight for freedom. The book compellingly shows how patriots like Samuel Adams, Patrick Henry, John Adams, Abigail Adams, Benjamin Rush, James Otis, Henry Knox, Paul Revere, John Hancock, Thomas Paine, George Washington, Martha Washington, and others, understood the conflict not merely as resistance to British overreach, but as a defense of God-given liberty against arbitrary, and often brutal, power.
A particularly striking theme throughout the book is the Founding Fathers’ deep reliance on Divine Providence. Metaxas highlights how many believed that human rights flowed not from kings or governments, but from God Himself, and that the outcome of the Revolution ultimately rested in His hands. Prayer, biblical conviction, and appeals to Providence are woven into the narrative, reminding readers that the Founders often saw themselves as instruments in a larger story unfolding under God’s sovereign care. Indeed, Providence often revealed His hand in altering the weather and divinely guiding the patriots as they fought the British forces. The result is a sweeping and engaging account of America’s birth that frames the Revolution as both a historical and spiritual struggle, fought with courage, conviction, virtue, and faith.
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Reviewed in the United States on June 2, 2026
★★★★★ 5
A New Classic
Format: Kindle
I pre-ordered, so I was able to get the PDF and start reading early. Of all the books I own on the American Revolution, I think this one stands to become my "go to." It is encyclopedic in scope, but deftly avoids becoming a drudgery or a slog through a mire of textbook facts. Somehow, Eric has managed to include a great deal of detail without losing any of the drama. It’s the story that rises to the fore in a gripping tale that just happens to be true (and much more than that, is our heritage). All the trademark “Metaxas” hallmarks are here: judiciously placed humor and lightheartedness that never descends into silliness, seriousness that somehow never lacks in winsomeness, depth that never smothers. He manages to portray both the soberness that the subject deserves without losing the hopefulness that the story of the Revolution should inspire in us. And along the way, you’ll find yourself better understanding what “went down” and (just as importantly) what it means to you today. Excellent work. If you think another history book on the American Revolution cannot inspire you or keep your attention, this one is up to the challenge.
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Reviewed in the United States on June 2, 2026
★★★★★ 5
The History Teacher I Always Wished For
Format: Hardcover
With his trademark wit and his eye for those wonderfully absurd details that most historians tend to overlook, Metaxas drew me deeply into the story. I found myself laughing out loud one moment and profoundly moved the next. The tiny anecdotes and humorous asides were not distractions at all — they were precisely what made these towering figures suddenly feel human and unforgettable.
Eric makes these historical figures flesh-and-blood again. He brings both heroes and villains vividly to life in a way that makes the drama of America’s birth feel immediate, thrilling, and deeply personal.
In short, Eric Metaxas is the history teacher I always wished I had. At 600 pages, this book reads with the ease and momentum of a paperback novel, yet carries extraordinary depth and scholarship. In the end, I came away with a renewed love for American history and a deeper gratitude for the souls who made this great country possible.
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Reviewed in the United States on June 2, 2026
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