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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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4.3 ★★★★★
Based on 185 reviews
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Product Reviews
★★★★★ 5
Westies Fav
Size: Small, Pattern Name: Pack of 1
Best Westie ball ever, my dog loves this ball. Exactly her size! 3" tennis balls are too big & she rips the fur off. this silicone-ish ball has no fur & is 2&1/2 "
We have 2" balls from her electronic ball laucher, but she still loves this glow in the dark best!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 4, 2026
★★★★★ 5
Best dog ball in the world!
Size: Large, Pattern Name: Pack of 1, Size: Large, Pattern Name: Pack of 1
My dog is singularly obsessed with playing ball. He would drop a steak to chase a ball. (He'd return to the steak, but not without the retrieved ball. He's not an idiot.) He makes dangerous leaps, crashes into trees, and runs fast enough to catch thrown balls before they even hit the ground. Since he doesn't care about his own wellbeing, it's my job to protect him as much as I can. And a lot of balls are hard enough to cause him pain or dental damage when he mistimes a catch. Enter the ChuckIt Glow Ball!
The minute Cowboy got his first Glow Ball, he no longer had any interest in the dozens of tennis balls scattered around the house and yard. If I took the Glow Ball away and instead threw a tennis ball, Cowboy would run to retrieve the ball, then return without it and begin looking all over the house for the Glow Ball. The Glow Ball is tough enough to survive Cowboy's big, sharp chompers, yet it's soft enough to do no harm when it hits him in the face. The glow feature is a huge bonus, too. Cowboy gets distracted by chipmunks and squirrels when we're playing, so he sometimes can't remember where he dropped the ball to pursue the interloper. If the ball was exposed to enough light before it was misplaced, it will be easy to find once the sun goes down. (Otherwise, Cowboy always finds it eventually.) When I'm feeling nocturnal, I'll put the ball under a light for a few minutes to activate the electrons, then treat my boy to a middle-of-the-night game of fetch.
I was so happy to see these on sale, I bought a lifetime supply for my pantry. The size guide seems accurate. ChuckIt recommended the large size for a dog of Cowboy's weight and that size is perfect. Highly recommended product.
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Reviewed in the United States on October 14, 2024
★★★★★ 5
Perfect for Blind Dogs, Fun and Durable
Size: 2-pack, Style: Duo
My blind dog loves these Chuckit! sniff balls. She can easily sniff them out and plays fetch with them for 20 minutes a day, even running up and down the stairs.
The balls are durable, bounce well, and the bacon and peanut butter scent keeps her engaged. I do wish Chuckit! would bring back the version with a tone so we could play fetch outside more easily.
Overall, a fantastic toy for dogs that rely on scent and a fun way to keep them active.
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Reviewed in the United States on February 14, 2026
★★★★★ 5
My Golden Loves
Size: 2-pack, Style: Duo
My 8 month old Golden Retriever loves these. It is also one of the few products she cannot destroy. They smell as described and she loves to chew them and play fetch when. She can chew these easily without destroying them. Overall good value for the money for my Golden
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Reviewed in the United States on January 2, 2026
★★★★★ 5
My dogs are obsessed with this ball
Size: 2-pack, Style: Duo
Great quality! I have 2 boxers that love it and the ball is still intact
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 31, 2026
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