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Description
Mouse VDAC1 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 utilizes 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 voltage-dependent anion-selective channel 1 (VDAC1) capture antibody. 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 voltage-dependent anion-selective channel 1 (VDAC1) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Voltage-dependent anion-selective channel 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Voltage-dependent anion channel protein 1, also known as VDAC1, is encoded by the VDAC1 gene. It is a beta-barrel protein that forms ion channels in the outer mitochondrial membrane (OMM) and the cell membrane. In the OMM, it allows ATP to diffuse from the mitochondria into the cytoplasm, while in the cell membrane, it participates in volume regulation. In all eukaryotic cells, mitochondria are responsible for synthesizing ATP and other metabolites required for cell survival. Therefore, VDAC1 allows communication between mitochondria and the cell, mediating the balance between cellular metabolism and cell death. | |||||||||||||||||||||||||||||||||
| 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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4.9 ★★★★★
Based on 1618 reviews
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Product Reviews
★★★★★ 5
One of the best Star Wars comic series of all time.
Format: Kindle
Crimson Empire was the first Star Wars comic that I read. Stories like these are what kept Star Wars alive and fresh during the nearly 20 years between films.
I love the stories about those minor characters that you see in the films. When you saw the Crimson Guard with the Emperor for the first time, you knew there was something fascinating about these guardians of the most powerful being in the universe. This story delves into some of those characters.
The art is fantastic, sort of that stylized 90's colorful art. Awesome battle scenes. It really captures the imagination.
Great story, give it a try. I highly recommend getting a paper copy of this trade.
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Reviewed in the United States on August 10, 2015
★★★★★ 5
Top Notch!
Format: Hardcover
A lot of the time when your reading SW books your imagination doesn't quite fill in the gaps that the author leaves. A lot of SW books have little to no detail because they are movie or game based. This is where the top notch illiustrations took this story line to a whole new level! Richardson does a great job on developping the main character. I was very impressed with the quality of the story and blown away with the illustrations. Great read for all!
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Reviewed in the United States on September 23, 2013
★★★★★ 5
Great saga!
Format: Kindle
I love this series! It is beautifully painted, has good dialogue, and has a very compelling narrative. I cannot help but like the protagonist, Kir Kanos. He is brave, unselfish, noble, determined, and deadly. Even if his primary goal is to exact vengeance for the killing of the truly evil Palpatine, I cannot help but cheer for him. This is truly a compelling read that refuses to be put down.
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Reviewed in the United States on October 21, 2016
★★★★★ 3
Just an OK read
Format: Kindle
I was not aware when I purchased this that it was a Graphic Novel, I really wish that in the blurb about these products it was made very clear that you are buying a Graphic Novel. I like reading, not viewing the written word. Since I got this via Kindle there was no shipping issues. Maybe the Graphic Novels should have their own category, so that we don't end up with things we don't want.
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Reviewed in the United States on July 25, 2015
★★★★★ 4
"Thus die all traitors."
Format: Hardcover
At a grand 504 pages, this big book covers the Crimson Empire series in its entirety. Containing the first, second, and third mini-series as well as Bounty Hunters: Kenix Kil, Dark Horse Extra #21-24 "Hard Currency", and Dark Horse Presents #1 "Third Time Pays for All". While some of these stories truly pale in comparison to the original series, they all still form a big story that is collected in this book. Slightly smaller than a regular TPB, this hardcover edition looks nice with a dustjacket (although mine was very off-center) but utilizes a glued binding on this thick book so you lose a bit to gutter loss.
The first story in the book is the classic Crimson Empire series. The six-issue series is collected here in full with a truly timeless story by Randy Stradley and Mike Richardson. Paul Gulacy did the awesome art within the issues. The writing and art work well together with the vibration of the blades to the movie-like, choreographed 12-page fight scene between Kanos and Jax at the end. A truly epic tale with lots of action and mystery that made you feel like you were watching another Star Wars movie but condensed into six issues of a comic book! This alone is worth the purchase price but you get even more stories after this!
Bounty Hunters: Kenix Kil follows the Crimson Empire in a tale following Kir Kanos after the end of the Crimson Empire series and was the third issue in the Bounty Hunters series. Kir becomes the bounty hunter Kenix Kil to move through a bounty hunter-filled planet and get what he needs and get out alive! Javier Saltares did the penciling while Randy Stradley reprised his role for the story. The story's short but tells a bit more about Kir and his journey. The drawings, while not as good as the first series, look good enough to get the story across.
Crimson Empire II: Council of Blood is next directly following the first series as Kir Kanos, as Kenix Kil, continues his quest to destroy what's left of the traitorous Imperial leaders. However, the return of an old friend side-tracks his quest and brings him to an even bigger journey! The old writing team of Mike Richardson and Randy Stradley return in this story as well as the original artist Paul Gulacy. The art's great and the story, though a bit dense, works well. There isn't quite as much action this time around but the story's just as good. The Zanzibar creatures are one of the creepiest things you'll ever see in a Star Wars comic, too!
Next up is the very short four-part comic entitled Hard Currency that appeared in Dark Horse Extra #21-24. The comic is written by Randy Stradley so you know the writing's done well but the art is by Isaas Buckminister Owens and is one God-awful mess. The characters are horribly out of proportion and it looks extremely cartoony. It's very, very short with only a few pages but even if you get past the art, the comic reads like a calendar with the book turned on its side. So, the whole process of reading this out of a 500+ page book is just annoying. I know they probably couldn't print it any other way but it's still inconvenient. However, what you get is a neat story wrapping up the fate of a character that has ran through the first two series and a bit more about Kir's alter ego Kenix Kil. Unlisted, the book appears to start with the third main series but actually contains an 8-page prequel comic that originally appeared in Dark Horse Presents #1 entitled The Third Time Pays for All. The writing has Randy Stradley again and, thankfully, Paul Gulacy on art duty (although his other works here were better). Once again, a short glimpse into the life of (a newly outfitted) Kenix Kil on a bounty-hunting mission while he reminisces about his past run-ins with Mirith Sinn.
Mike, Randy and Paul continue their work with the Crimson Empire III: Empire Lost where Kir Kanos rejoins Mirith Sinn one last time to thwart an Imperial thug from destroying the New Republic and the New Empire in one fell swoop! Leia, Luke, Han, and Chewie appear in this tale as well as Boba Fett to round out a classic cast. The art's great, once again, and the writing, while probably my least favorite of the series, is still pretty good with an epic fight between Kir and Devian. At the end of the book, we get the Crimson Empire Handbook entries on some of the characters as well as a few more covers to gawk at. While this hardcover book looks really nice, Dark Horse still fails to make a truly great edition for this series through the book itself. The contents are great but the small size and lack of comic covers are disappointing. Sadly, that's just how Dark Horse releases their hardcovers and TPBs. But, if you're looking to read the Crimson Empire books, this is the one to get!
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Reviewed in the United States on August 22, 2013