Pay in installments of $46.03 with
,
and
Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Sep 4 - Sep 9
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
Description
Human PDI ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. 4. 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 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 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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. |
|||||||||||||||||||||||||||||||||
| 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 capture antibody against protein disulfide isomerase (PDI). 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 protein disulfide isomerase (PDI) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Protein Disulfide Isomerase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
|
|||||||||||||||||||||||||||||||||
| Background | Protein disulfide isomerase (PDI) is an enzyme found in the endoplasmic reticulum (ER) of eukaryotes and the periplasm of bacteria. It catalyzes the formation and cleavage of disulfide bonds between cysteine residues in proteins during protein folding. This allows proteins to quickly find the correct arrangement of disulfide bonds in a fully folded state, thus catalyzing protein folding. | |||||||||||||||||||||||||||||||||
| 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. |
|||||||||||||||||||||||||||||||||
| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
Shipping Notes
- Free Standard Shipping on $100+ Orders to the USA.
- Except Preorder products are shipped in 48 hours.
- Delivery to the USA:
- Standard Shipping : 3-10 business days
- If time is of the essence, please consider selecting expedited delivery for faster service.
Exchange/Return Notes
- We offer a 30-day return/exchange service after receiving.
- Final sale items are not eligible for returns or exchanges.
- To process your return/exchange, please contact us at [email protected]
- Please click here for more details>>> Return & Exchange Policy
4.5 ★★★★★
Based on 8 reviews
Sort
Product Reviews
★★★★★ 4
Interesting and clear
Format: Kindle
The book reviews the history of the law that protects interactive websites from prosecution due to defamation. The discussion is interesting and straightforward, but there are too many unimportant details in some places.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 2, 2022
★★★★★ 5
The most well researched book about Section 230 and free speech on the Internet
Format: Hardcover
Many are calling for regulation of the tech industry. This book does an excellent job of outlining the “it’s not that simple” aspect of the debate. Some say big tech is doing too much to silence legitimate conversations and others say they are not doing enough to combat hate speech and false information.
Regardless of which side you fall on it is important to know the history and this book is a well written account. Professor Kosseff does an excellent job of both presenting the history of Section 230, but also mixing in his well reasoned opinions about how we should proceed. Most importantly posing the question of if you don’t want big tech regulating the internet, you better think long and hard about who you do want, and what that will look like.
This should be a mandatory read for any tech reporter who even utters the words “Section 230”. Thanks for the information.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 26, 2019
★★★★★ 5
The book that might save the internet
Format: Hardcover
The level of misunderstanding about Section 230 in media and government is astounding. All they have to do is read Kosseff's book to clear up their misunderstanding. The New York Times made a colossal mistake confusing Section 230 with the First Amendment. I hear media people all the time thinking they can't moderate comments because of Section 230 (the opposite is true). I see politicians going after Section 230 because they think it is the root of the net's problems (they are dead wrong). Section 230 is our best protection of free expression and human dignity on the net, for as Kosseff clearly explains, it is intended to enable the public conversation (by not making platforms -- including news organizations -- liable for what others do on their sites) and it is also intended to empower platforms and media companies to moderate what people do on their sites to their own standards. The law is short and clear and is vital to the future of the net and the public conversation on it.
Kosseff puts forward an excellent history of the development of legislation and case law on free expression in books, movies, and other media. He talks with the creators of Section 230 and explains their goals and motives. He outlines the debate about the law.
That debate is now reaching a critical phase in both media and Washington. Thus Kosseff's book is a document of urgent importance. I wish some rich person would come along to buy every member of Congress and every news organization a copy so they don't keep screwing this up.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 5, 2019
★★★★★ 3
Please use a larger font
Format: Hardcover
Over 60 and sometimes have a difficult time with too small print? My advise .. either read it on an ebook or skip this one.
I read the first chapter despite the small font size that made it a little too difficult to read. While I found it informative and well worth reading I'm probably not going to read anymore and return it to the library. Mass market paperback books with small print I can understand. Hardback non-fiction books (which are generally read by an older crowd I suspect) make no sense to me.
5 stars for the content I read. 1 star (generously) for the font size.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 6, 2020
★★★★★ 5
Great review and analysis of a law that underpins the Internet
Format: Kindle
This book provides a detailed history of how section 230 if the Communications Decency Act came to be, its importance and its problems as the industry of the Internet matures. While it is ultimately a defense of that law, it is an eyes wide open look at it as well. It is not a prophet's telling of what to do, but instead an informative read that gives the reader an ability to assess what might be done to address the issues 230 raises.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 9, 2019