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Description
Human FUNDC1 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 frozen and thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 a FUN14 domain containing 1 (FUNDC1) 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 amount of FUN14 domain containing 1 (FUNDC1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human FUN14 domain containing 1ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | FUN14 domain-containing protein 1 (FUNDC1) is an integral mitochondrial outer membrane protein that plays a key role as a receptor for hypoxia-induced mitophagy. It is encoded by the FUNDC1 gene. Pathways involved include selective autophagy and mitophagy. An important homologue of this gene is FUNDC2. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.5 ★★★★★
Based on 15 reviews
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Product Reviews
★★★★★ 5
better than the series
Format: Kindle
The chemistry the characters the banter the amazing direction of the story all builds up to a crescendo that simple didn’t let go! This is the first of an epic series that simply let love in its many different ways and journey transformed lives in ways that one couldn’t imagine! Beautifully worded and crafted with a creative nack to entertain and really dig this universe! Well done Miss Kennedy, well done!
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Reviewed in the United States on May 29, 2026
★★★★★ 4
Not forced or rushed
Format: Hardcover
I really liked this book. Hannah & Garrett’s story starts off like a typical smart girl tutors the jock to help him get though a class in college. But their stories are traumatic and the things that they have both had to overcome make this book so much more than your typical tutor/jock story. Garrett is sweet, cocky, funny and helps bring Hannah out of her shell in a refreshing way. Hannah is not meek or mild or quiet, she is strong because of her past but still a little unsure. Together their chemistry is hot and their relationship just sort of naturally unfolds. I really loved that it didn’t seem forced or rushed, it just happened as it should. Garrett’s story in particular hit very close for me so maybe that is why I felt connected to these characters so much. I am really looking forward to reading the rest of the series.
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Reviewed in the United States on May 21, 2026
★★★★★ 5
The Hype Train Goes 🚂
Format: Kindle
4.5⭐️ | 4🌶️ | 4🎧
I genuinely enjoyed this book! It’s been sitting on my TBR for ages, and with all the buzz surrounding it lately, I finally decided to hop on the hype train. Thankfully, this one absolutely delivered.
What surprised me most was how much depth this story had. Beneath the humor, hockey, and romance are some pretty heavy topics, and I thought Elle Kennedy handled them with a lot of care. Hannah’s journey in particular felt incredibly authentic, and I appreciated how thoughtfully the book approached her past trauma. This story felt like a safe space for survivors, never minimizing what she experienced while still allowing her healing and growth to shine through.
Garrett and Hannah were such a fun couple to follow. Their banter was hilarious, their chemistry was undeniable, and their relationship developed in a way that kept me completely invested. I loved watching them challenge each other, support each other, and slowly become exactly what the other person needed. The romance was sweet, the spice delivered, and the emotional moments hit when they needed to.
As a hockey fan, I also loved the sports aspect of the story. Hockey is one of the few sports I actually watch, so those scenes were especially enjoyable for me. But more than anything, I adored the sense of found family throughout the book. The friendships and team dynamics added so much heart and made the world feel lived in and welcoming.
The audiobook was solid overall. Christian Fox and Lorelei Avalon did a good job bringing the characters to life. Christian Fox was a standout for me, while Lorelei Avalon occasionally sounded a little robotic during certain scenes. It wasn’t enough to take away from my enjoyment, but it was distracting at times.
Overall, I can definitely see why so many readers love this book. It’s funny, emotional, spicy, and full of heart. I’m excited to continue with the rest of the series and spend more time with this group of characters! 💕⛸️🏒
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Reviewed in the United States on June 4, 2026
★★★★★ 5
Dreamy Hockey Men & Steamy Romance
Format: Audiobook
Loved this book! This book came up as one of my Amazon similars. As for contemporary, they must have an intriguing plot to hold my interest. I normally prefer Paranormal, Dystopian or Sci-fi romance or romantic suspense.
The blurb sounded appealing, so I decided to give it a chance. This was my first hockey romance. After reading this book, I'm now a fan of the genre. This story had everything in it that I loved. An intense, slow-built romance. A cocky guy, who didn't really do relationships until he meets the girl who brings him to his knees. Friends to lovers with a few games in between. Spicy love scenes and jealous. Most importantly, a driving plot with intense drama that had the perfect balance. But it wasn't childish or over the top.
I was over the moon when I recently learned that this book is becoming an adaptation for on Amazon. May 13th can't come fast enough.
This book was so good, I decided to check out book #3 in the series, The Score. All 4 books in the Spin off series blurbs sounded interesting. After book #3, I immediately read all the books in the spin-off. Book #1, The Deal is tied with The Play in the spin-off series as my favorite with The Risk in the spin-off being a close 2nd..
Another thing I like about her series is that they all the books are stand-alones. You can also read the books out of order. Each book is a separate couple, so you get a new story each time, alongside following the other characters that were introduced in previous books
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Reviewed in the United States on May 19, 2026
★★★★★ 4
Good Start
Format: Kindle
This was the first book in the "Off-Campus" series. Hannah Wells, a focused and reserved music student, and Garrett Graham, the confident captain of the college hockey team whose academic struggles threaten his future. When their paths cross through an unexpected agreement, a friendship begins that gradually deepens into something more meaningful. I normally read YA but I found that I liked both of these characters I also liked the secondary characters as well. This was a pretty good read and I see why it's a good series on prime. This book was well written with no errors in grammar or spelling. I am looking forward to reading the next book in this series.
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Reviewed in the United States on May 16, 2026