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Description
Rat COL2 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 40 ng/mL). Then dilute to the following concentrations: 40 ng/mL, 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 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 40 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 20 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. |
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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 collagen type II (COL2) capture antibody. After incubation and washing, the sample 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 collagen type II (COL2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Collagen Type II ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Type II collagen (COL2) is the building block of articular cartilage and hyaline cartilage. It is composed of the α1 chain of the type II homocollagen. It accounts for 50% of all proteins in cartilage and 85-90% of the collagen in articular cartilage. Type II collagen forms fibrous structures. This fibrous collagen network enables cartilage to hold aggregates of proteoglycans and provides tensile strength to the tissue. | |||||||||||||||||||||||||||||||||
| 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 | 0.62-40 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.7 ★★★★★
Based on 23 reviews
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Product Reviews
★★★★★ 5
What is better? The overachieving 6 or underachieving 8?
Format: Hardcover
First, let me say that I thought that this was an excellent book and would recommend it to anyone who is at all interested in the college admissions process. Second, I was surprised at how many of the reviewers seemed shocked--shocked!--that applicants got bonus points for coming from minority backgrounds. Was this some kind of revelation? However one thing that surprised me a little bit is how--even moving beyond race entirely--the more advantages you have had in life, the more disadvantageous it will be for your admissions process. For example, I was unaware that having successful parents would be, in essence, held against you on the theory that more would be expected of you. While other reviewers have (jokingly?) said that they would advise their white kids not to check the "Caucasian" box, I might advise my (still very young) kids to say that their parents have been unemployed their whole life.
I suppose that the main issue which this whole process really boils down to is the following: As a college applicant, is it more important to succeed in life relative to the world around you (i.e. relative to your classmates, to others of your race, to others of your geographical area, to your own parents' life and accomplishments, etc.) or is it more important to succeed absolutely and not on a relative scale. This book clearly informs us that the answer is the former and not the latter. Whether that should be the answer is another question.
For example, say that a student's entire life could be distilled into 2 numbers each on a sliding scale from 1-10. The first number is simply your academic performance (grades, SAT's, course load, etc.) The second number is your background (race, economic circumstances, gender, etc.) In the case of Wesleyan, it seems clear to me that they would rather have a student whose first number was, say, a 6 if his or her second was a 2 (take Mig for example in Steinberg's book) than a student whose first number was an 8 if the second number was a 9 or 10 (take Tiffany Wang for example). Whether that is the right approach is certainly a legitimate issue for discusion and I'm not saying that it's not.
I suppose that one of the things that would be interesting to know (even though one never really can know of course) is whether those numbers will change in the future. For example, if one were to know that Mig would always be a 6 and Tiffany would always be an 8, would that change the analysis as to which is the right approach? I suspect that part of the reason that a school like Wesleyan would favor the overachieving 6 over the underachieving 8 is due to the hope or expectation that those trends will continue in the future and that one day the 6 will actually be ahead of the 8. And maybe that's the way it works. Who knows.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 28, 2003
★★★★★ 5
You will find out how a selective private college evaluate and admit students
Format: Paperback
I'm a high school counselor and college advisor. Fifteen years ago when I started my college counseling position, I struggled to understand or explain to students and their parents how a selective private college evaluate and admit students. It was this book that helped me understand the essence of selective private college admissions. Compared to other dry theory books, this book tells the admissions practice as stories that are easy to read, understand, and associate with. I highly recommend this book to students, parents, and new counselors.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 5, 2024
★★★★★ 3
Who edited this mess?!?!?!?
Format: Kindle
This is a very interesting work of nonfiction. I found it intriguing and read it very quickly. I actually got invested in these students and their stories and their journey to get admitted to the college that was right for them. BUT, and this is a big but, this book is so poorly edited, it is disgraceful! If a person were reading this for research purposes, and it could be useful for just that, good luck to them. The dates are all over the place. At one point, the kids are being considered for the class of 2004, then it makes a reference to the current year as 2000, then it reverts back to 2004 for a long while, then it mentions how the kids--currently at their various chosen colleges--reacted to the events of 9/11/01. What the hell? It's very confusing. It makes it very difficult to keep things in context.
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Reviewed in the United States on August 8, 2013
★★★★★ 5
Abundant information but needs update
Format: Kindle
I struggled a little when try to decide how many star to give. It's an excellent book and very informative. It feels like I was not reading a college admission advisory book, which very often are dry and mechanical. It feels like reading stories of students and AOs, I got attached and involved emotionally, and really felt for them. But there were abundant information in the book, I was able to use the book to answer most my questions. If you need a list of YESs, Nos, Warnings..., then this is not the book for you. If you need to know what happens during admission process, then this is an excellent book for you. Based on the information in this book, I was able to extract my own conclusions.
The reason I hesitated if I should rate this book as a five stars book is how long ago it was written. A lot has changed. Many aspects stayed same, but many aspects changed. It will be misleading if this is the only guide book you are using. Wish everyone a great college application season.
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Reviewed in the United States on September 12, 2015
★★★★★ 4
An intimidating read
Format: Paperback
This is a scary book to read if you are anyone, but most especially a Parent of a college bound junior or senior. The writer, Jacques Steinberg, a NY Times Reporter writes beautifully and captures the college application process in an authentic way, without overdramatization. His style and sensibility reminds me of Jon Kraukeur. I was prinicipally struck by the heroic Sisyphean efforts of the admissions office and the immense gap between public school and private school admissions counseling at the high school level, a gap which is even greater today. This is also a very good description of Wesleyan University -- what it is not so good at, and this is no fault of the author who tries to make this point and the related one that follows , but the best approach for the applicant is to find a college where he and she can thrive and trust the process, the admissions folks are very dedicated to bringing in students who can add value and to whom they can add value -- more to the point when a good fit is made, all the effort and anxiety is well worthwhile to the school and the applicant. good luck and best wishes to all of you who are college bound.
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Reviewed in the United States on December 9, 2012
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