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Description
Rat CAV1 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: 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: 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. 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. 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. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, 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, avoiding repeated freeze-thaw cycles. 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 caveolin 1 (CAV1) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of caveolin 1 (CAV1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Caveolin 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Caveolin (CAV1) is a protein encoded by the CAV1 gene. The scaffolding protein encoded by this gene is a major component of the plasma membrane of caveolae found in most cell types. This protein links integrin subunits to the tyrosine kinase FYN, an initial step in the integrin-Ras-ERK pathway and promotes cell cycle progression. It has been shown to interact with heterotrimeric G proteins, Src tyrosine kinases (Src, Lyn) and H-Ras, cholesterol, TGF-beta receptor 1, endothelial NOS, androgen receptor, amyloid precursor protein, connexins, and α1. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.7 ★★★★★
Based on 7 reviews
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Product Reviews
★★★★★ 5
What a Great Ride
Format: Kindle
This book was a fun ride from start to finish. While not fully LitRPG, but sorta' LitRPG light based on the system that's set up, the world building makes sense and fits the story very well.
The real shining star was the character development. I felt like I knew the characters after a few lines and as the story developed, casual comments helped build a deeper web of interrelationships.
Also loved the blurbs at the start of the chapters - thought they really added to the world building and backstory in a fun way.
Can't wait to read more about Cat!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 3, 2021
★★★★★ 5
Wonderful
Format: Kindle
This was everything I was hoping for and more. Great world building, awesome characters and a mythology that just left you wanting more. Little snippets at the start of each chapter that hints at the history of the world at large while also slightly framing the content of that chapter? Spot on. My one issue, and I believe others have pointed it out, is the fact that the text is grey and not white, which can make it harder to read when you’re, say, outside. But that’s a formatting thing and has no bearing on the contents. I’ll be waiting in line to get the sequel as soon as it appears! Oh, since Amazon can be iffy about it, the LGBTQ+ tag is accurate!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 28, 2021
★★★★★ 4
I love this author's storytelling!
Format: Kindle
This is my fourth series by Ravensdagger, and I am truly addicted to the masterful storytelling. This series is just as entertaining and full of elaborate world and character building as I have come to expect. That said, I docked a star for the first time ever with this author because this is the first of the author's series I have experienced that contains the rampant and horrifyingly overused profanity of so many authors in this genre. Honestly, it was so disappointing and so prolific in this book that if it hadn't been just an awesome story full of so much depth and entertainment, I would have docked one or two more stars. We simply don't need those virus words constantly infecting, narrowing, and eventually taking over our much broader vocabularies as they do so very often. But the fact that I left 4 stars really does speak to how much I enjoyed the book other than that. I can't wait to start the next one, and I hope it will be as refreshingly free of profanities as this rest of this Ravensdagger's excellent body of work.
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Reviewed in the United States on March 12, 2024
★★★★★ 5
A Must-Read for College (and Even High School) Students
Format: Paperback
First of all, this book really cuts to the point. If you are skilled with skim-reading, you can get this book in about 2 1/2 to 3 hours.
Second, while these strategies are currently untested by me, they seem really practical. Rather than saying "study" or "write paper" or "take test", Dr. Newport lays down specific, useable guidelines for all areas that are intended to cut down on time spent on schoolwork. This helps you with more time to have fun, do extracurricular activities, or work 60 hours a week (like I will soon be doing).
Third, this is THE anti-procrastination book, although I don't get the concept of starting term papers a month before they're due as the examples did. The whole concept behind this book is making sure you've got as little work going as possible that takes as little time as possible. Doing things upfront will ease your load later when the assignments and papers start rolling in and out.
There are only a couple of issues I have with the book.
1) Dr. Newport almost makes the use of personal laptops compulsory in college. I get why it would be such an asset, but for some of us it is more of a liability than an asset. I really believe one can get through college without a laptop at a much smaller personal expense than the costs of buying a laptop/accessories, printer, ink, etc. If you already bought one, that's one thing, but if you haven't, why do you need one? Just use pen/paper and the computer labs' computers and printers. With the printers, odds are you're paying the same amount for 1000 pages that you would for toner (paper is free with the way they price it), and you don't have to drop the cost upfront, but pay as you need the printer. Financially, you're better off skipping the personal printer.
2) He didn't discuss what format to purchase textbooks, but considering he was so insistent on portability, I would probably haphazard a guess that a hard copy is the only way to go. Sure it may be expensive, but what if your Kindle or iPad gets stolen?
Overall, highly, highly recommended. Get this for your child as an early (or very early) graduation present. Teaching them these things will save them a lot of headaches later on.
One last note: This book is not for the lazy. If you aren't willing to do the work, don't expect this book to help. However, if you're someone like me who's sick of lazy and wants to convert, this book IS for you!
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Reviewed in the United States on December 18, 2012
★★★★★ 5
One more crappy college study guide
Format: Paperback
When I was taking 4 classes at my local community college, I tried to follow the part of the book that said just to learn math by examples. I ended having to drop the class because at first I was actually ahead of the professor. Then I tried this stupid book's technique of learning by examples and I ended up falling behind. The same can be said of "What smart students know" it's actually a good technique (smart students), but you end up spending hours thinking up questions. Questions can be a part of the process, but it can't be the whole process. I just bought an online course than is way better than both of these combined. It's so good. The point I'm at in the course you can memorize up to 50 items in reverse and forward. You can also tell which place in the list. An example could be
The order of my hallway:
Dog = front door
Cat = wall
Duck = light switch
Printer = holder
Hitler = chair
MAO = rug
Goku = ceiling light
Kurrin = mirror
Gohan = heater
Frieza = painting
You would then connect / visualize each word / picture with a part of your house. The rule is to visualize each for only 6 seconds while recalling can only take 6 seconds. If you miss one you have to do the six second process again until you make no mistakes.
P.S. I'm not telling anyone the name of the course or where to buy it. I want to be ahead of all other students.
Edit: I'm back in college taking two college level courses. While I don't agree with everything in this book (I have to re-read it) I agree with 80-85% of the methods used in this book that I remember. Such as not doing the reading he was right. I haven't done the reading in my psychology 101 class and have only focused on taking / memorizing / learning the notes and while I haven't gotten a grade yet, I actually feel I'm learning enough and today in class I note notes from a documentary we watched. The professor said to do the reading for chapter 2 of the textbook. I'm not even going to do it. I'm going to just take notes in class then go over the reading. I did this for my 1st set of notes and I didn't understand something in the lecture notes. I ended up re-reading that / part / example [it was on the scientific method].
I remember from taking my developmental math class that the professor was actually doing examples (even though I couldn't figure out from what part of the book they were from and / or I couldn't write every example like he said) If I had just asked questions and either watched videos on Youtube I would've probably remembered the material from that class.
I'll do another edit once I'm done with classes. I'm also going to write / provide a list of books that I think are good for learning and have good advice for college.
Edit: I ended up trying to do the advice from this book and if I didn't look up the answers for the test for my psychology class I wouldn't be in college today. I'm not saying that all of this book is bad but I went from thinking that 85% of it is good to now thinking that 50% of this book is good. The other half is crap. Such as the technique for Q/E/C he doesn't give any examples or enough examples. The other advice like making your own problem sets I wouldn't have thought of. Even though this is an all right technique your wasting time because you can just practice doing other peoples problems (like those online or from a professor) or do Bullet point concept from another book I read (I'm going to provide a list once I'm done with this review). The one technique that is very good is the lecture technique he mentions in the book. For some reason, this technique helps with putting material into memory. His section on writing papers is also very good. His explanation on how to prepare for and take exams is also good. Also, he should have made the book more visual and put actual writing from actual students other than that it's an all right book.
Here's the list of books I've read (so far) that will most likely get you A's
College Rules
https://www.amazon.com/College-Rules-4th-Survive-Succeed/dp/1607748525/ref=sr_1_1?s=books&ie=UTF8&qid=1530133408&sr=1-1&keywords=college+rules
Guaranteed 4.0
https://www.amazon.com/Guaranteed-Follow-3-Step-Plan-Dont/dp/0974264806/ref=sr_1_1?s=books&ie=UTF8&qid=1530134451&sr=1-1&keywords=guranteed+4.0
How to study in college
https://www.amazon.com/How-Study-College-Walter-Pauk/dp/1133960782/ref=sr_1_4?s=books&ie=UTF8&qid=1530134633&sr=1-4&keywords=how+to+study+in+college
The only book I've read from cover to cover is GT 4.0 it's probably the best in my opinion because the system in this book will get you A's. The only reason last semester I didn't get A's was because I didn't follow the system to the t. If you follow the GT 4.0 system to a t and don't get all A's she'll give you $100.
Hope this helps.
Edit 2020:
From what I remember reading this book, this applies to college level courses and even at the same time I tried using his question / evidence technique while taking a psy101 class and couldn't do it. Also he's right to a very large degree about living off of lecture notes. Living off of the lecture notes IMHO can only be done if it's a science science class or a college level math class. I'm currently taking a pre-req class and the professor explains everything. Along with the Guaranteed 4.0 technique this is a pretty decent book.
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Reviewed in the United States on January 24, 2012