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Description
Rat ND1 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 NADH-ubiquinone oxidoreductase chain 1 (ND1) capture antibodies. After incubation and washing, the assay 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 amount of NADH-ubiquinone oxidoreductase chain 1 (ND1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat NADH-ubiquinone oxidoreductase chain 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | NADH dehydrogenase 1 (NADHD1), also known as ND1 or MT-ND1, is an enzyme encoded by the MT-ND1 gene. This protein is a subunit of NADH dehydrogenase, located on the inner mitochondrial membrane and the largest of the five complexes in the electron transport chain. It is a subunit of complex I of the respiratory chain and is considered to be part of the minimal assembly of core proteins required to catalyze the dehydrogenation of NADH and electron transfer to ubiquinone (coenzyme Q10). | |||||||||||||||||||||||||||||||||
| 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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4.1 ★★★★★
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Product Reviews
★★★★★ 4
An inspiring and engaging tale.
Format: Kindle
A Place for Us is a heart-felt story of love, redemption and second chances.
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Reviewed in the United States on August 12, 2025
★★★★★ 5
A Place for Us
Format: Kindle
Oh! 15 gold stars for this one! This book is so poignant and so very brave and authentic! This book is a permission for us to choose what we will and will not accept in our relationships. This book is more than hope, this book is determination! Pure driven determination to not settle, but to go to the mat to have our life the way we want we want it. I cannot over recommend this book. Thank you, Patricia Grayhall! Keep writing!
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Reviewed in the United States on June 8, 2025
★★★★★ 5
Finding a Home
Format: Paperback
A Place for us is a beautifully crafted and deeply moving love story about two women--one American, one British--who first meet in their youth and reconnect decades later. At its core, the novel portrays their struggle to navigate restrictive immigration laws in pursuit of a life together. Their journey highlights the urgent need to protect and legalize same-sex marriage, especially at a time when such rights feel increasingly precarious. A timely and relevant read.
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Reviewed in the United States on August 14, 2025
★★★★★ 5
So good
Format: Paperback
RATING : ⭐️⭐️⭐️⭐️⭐️ SPICE: 🔥🔥 Hot damn Hannah did it again .if you want to read more sapphic romance, this one will absolutely blow your mind! Bryce and Daisy were everything I wanted and more! Fake dating will always be the superior trope, but when it’s forced proximity grumpy x sunshine and the grumpy one is secretly head over heels for the sunshine one, well I’m a damn puddle on the floor! I still can’t decide if I loved Bryce or Daisy more, but I do. Know this might just be my favorite book Hannah has ever written. ☀️ Fake Dating ☀️ Grumpy x Sunshine ☀️ Forced Proximity / Roommates ☀️.Tattoo Artist / Teacher ☀️ So much pining I love Bryce so much. She is a total hard shell but she has such a gooey inside and she showed it so well with Daisy. This whole setup was fantastic, Daisy needs a place to stay so Bryce gets volunteered to give up her spare room. Daisy think Bryce doesn’t like her, but then you get to Bryce’s POV and you realize how down bad she has it for Daisy. Then Daisy proposes the fake date and OMG the tension. The longing. The secret tattoos. I loved every bit of these two figuring each other out. Watching Bryce absolutely melt for Daisy was just so so good. This may be a slow burn, but when it burns…. It’s an inferno. And the tattoo parlor scene is imbedded in my brain forever. Just everything about this made me happy and I will forever love Cherry Peak, these characters and this series! Now bring on Darren!
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Reviewed in the United States on January 10, 2025
★★★★★ 5
Love them!
Format: Paperback
“Calling Daisy Mitchell mine is a dream that I never planned on coming true but always hoped would.”
Gahhh my first FF and I’m hooked! Thank you Hannah!
Daisy and Bryce are as opposites as can be, one is pure sunshine and the other is as icy as can be but they couldn’t be more perfect for each other!
I love the slow burn with all the sexual tension as they pretend to date while living together. Love the way they take care of each other, the love and admiration so evident in the way they touch and look at each other. Their nicknames for each other is just so cute and perfect! I love these two so much!
Such a great addition to the Cherry Peak series! We get more of Johnny, who happens to be Daisy’s twin and I just can’t get enough of him either, he’s just so fun. And of course we get to see more of the rest of gang. Love these found family and I can’t wait for more!
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Reviewed in the United States on January 19, 2025