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Description
Rat t-PA 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 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. |
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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 capture antibody against Plasminogen Activator, Tissue (t-PA). 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 Plasminogen Activator, Tissue (t-PA) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Plasminogen Activator, Tissue ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Tissue-type plasminogen activator (t-PA), also known as PLAT, is a protein involved in blood clot breakdown. It is a serine protease found in endothelial cells, the cells lining blood vessels. As an enzyme, it catalyzes the conversion of plasminogen to plasmin, the primary enzyme responsible for clot breakdown. tPA can be produced using recombinant biotechnology; tPA produced in this way is called recombinant tissue plasmapoietin activator (rtPA). Specific rtPAs include alteplase, replicon, and teneplase. They are clinically used to treat embolic or thrombotic strokes. Their use is contraindicated in hemorrhagic stroke and head trauma. In cases of toxicity, the antidote for tPA is aminocaproic acid. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.0 ★★★★★
Based on 15 reviews
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Product Reviews
★★★★★ 5
High Quality Rolling Chair
Color: Black/Silver
My son LOVES this desk chair. He has a drafting table to draw and do schoolwork on, and this provides him a comfortable chair to work in. It is sturdy and easy to clean. It does not fold, but it does roll easily. It was VERY EASY to put together. We were also pleased with the price. He will be able to use it for years to come.
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Reviewed in the United States on January 2, 2026
★★★★★ 5
The Greatest Chair I've Ever Sat In
Color: Black/Silver, Color: Black/Silver
When I was a boy, my first office chair that I used for PC and laptop work growing up was this chair. This was back before 2010 so it likely wasn't under the Amazon Basics brand but it was the exact same chair design. I've used that chair my entire life really and only had to buy this chair as a replacement as the leather started peeling significantly on my old chair after over 20 years of use. Even though the leather was heavily peeled, my old chair was still extremely comfortable and I would've kept on using it had I not gotten annoyed with having to pick up leather bits and pieces everyday from the peeling. I bought this chair as a replacement and the comfort is still as I remembered. No lower back pain, no butt numbness, no arm numbness, truly such a masterpiece of a chair where it lets you focus on the task at hand instead of focusing on the chair. Before I rebought this chair, I've bought & tried the Secretlab Titan Evo Nanogen, Furmax executive office chair, Neo executive office chair, RS high back gaming chair w/ flip-up armrests, Bestfair high-back office chair, FDW executive office chair, Herman Miller Aeron, and OFIKA executive office chair in search of a potential "upgrade" - every single one of these other chairs was inferior to the Amazon Basics high back office chair in one way or the other. The only thing I will note is that the pneumatic gas cylinder of the Amazon Basics office chair seems to be different from my original chair - it goes higher and has a higher minimum seat height. If you would like to perfectly replicate the original lower seat depth you will have to buy the S6124 extra short office chair gas lift cylinder from ChairPartsOnline - my only nitpick about such a phenomenal chair. In my attached picture I have the Amazon Basics chair w/ S6124 gas cylinder on the right next to the Furmax high-back office chair which although inferior is what I consider to be the 2nd best chair if for some reason you cannot get the Amazon Basics high-back office chair. To sum it up - I HIGHLY RECOMMEND THIS CHAIR!!!
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Reviewed in the United States on December 2, 2025
★★★★★ 4
Comfortable Office Chair
Color: Black/Silver
I have had several office chairs here in my office. Even though this office chair was a bit difficult to assemble, it appears to be a quality item. I was NOT impressed with the instructions - at all. More specific instructions would have been nice. Only time will tell if this was a good purchase and this chair is a good value. Not a bad price for the item as well.
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Reviewed in the United States on May 15, 2026
★★★★★ 5
All-around good chair, suitable for me at 6'6", comfortable and no issues
Pros:
The assembly is straightforward and doesn't require much strength; I would expect my mother to have no issue putting this chair together. If you're elderly or disabled, a relative or helper should have an easy and quick time of putting it together, and there is very little chance of making a mistake during assembly.
Sturdy at and slightly above the recommended weight limit. I peaked around 256 or 258lbs while using this chair, and it didn't show any signs of complaint then, and it hasn't shown signs of wear since.
Really comfortable. I'm an eight-hours-a-day user, almost without exception, and I have no complaints.
The leather holds up very well. I don't expect this to be true forever, but after approximately 1800 hours of use, there is no tear or warning sign.
It leans back super stably. That's the most important aspect for me.
It's tall enough to support my 6'6" frame. My back rests comfortably, so the odds are yours will fit, too. For the scary few who tower over me, and so you can search it easily, here's a height guide:
6'3" 6'4" 6'5" - The chair is good for your heights
6'7" - You should also be well served, unless your height is mostly in your torso.
6'8" - You're at the cusp. I think you will be comfortable in this chair, and alternatives that are built for our (mostly your) height cost so much more. I'd give it a try. I wish you luck.
6'9" 6'10" 6'11" - This is written for 6'9. For 6'10, 6'11, scale the significance of the caution. I think this chair will prove uncomfortable for you or encourage bad posture. I'd encourage you to look elsewhere.
Cons:
This is well covered in other reviews: the caps that cover up the screws on the side of the arms are poorly designed. One falls off every day or two. It's a minor inconvenience, but it's optional. You can toss the caps in the trash. The screws are not exposed. They sit deep within the plastic of the arm. Personally, I just push the covers back on when I notice they fell.
The armrests could be better for me, I think. I'm not sure exactly what my complaint is. I think they are a little too narrow for my liking, but I'm 6'6" with broad shoulders and long arms and unusual joints. The spacing seems fine for a typical user.
Notes:
I only sit leaning the chair back or sitting up straight at the edge of my seat. Some aspects of this review may not translate perfectly to locked-in-place, sitting back, upright usage.
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Reviewed in the United States on July 8, 2018
★★★★★ 5
Awesome!
Update: March 2022 - Funny reading my previous review now. All the cons I mentioned before, I literally don’t even notice at all. In fact, I like the chair I got so much I just got another in white. My boyfriend is using the old one and I needed one for my home office. And sadly, my cats did scratch up the black chair a little (the cats live outside now) but the chair has otherwise stood up to use perfectly. Other than the scratches, is basically like new. I do wish I knew of a good repair kit cause I would like to fix up the scratches. The chair is just extremely comfortable and supportive which I need even more these days for my home business. A very good purchase!!
I just received my chair 2 days after ordering it. It took me about 20 minutes to put it together which was very easy. I am sitting in it now typing this review.
Pros:
-Very attractive. I ordered the black color and it appears just as in the pictures.
-Slides smoothly over my linoleum floors.
-Seat is soft and comfortable, hopefully it will stay that way over time, but too soon to tell.
-Very easy to assemble.
-The seat is good size for the average American and seems comparable in width to the chairs I use at work.
Cons:
-The chair does not lower as much as I wanted. I like my legs at a 90 degree angle when sitting. It is close with my shoes on, but about 2 inches to high. If I had known this, I very likely would not have ordered this chair as its one of my most important features for sitting comfortably. I am 5'6”. I hope that with a little wear the cushion will compress a bit, and if not I may get a small foot stool to solve the problem.
-As others have described, this chair pretty much doesn't recline in any appreciable way even at the loosest setting. Basically to stay reclined you have to keep your legs pushing back the whole time which eliminates the point of reclining. I have read from others that over time, this loosens up. This feature is not very important to me, but may be to others.
-The seat depth is about 2 inches shorter than I would like.
Overall, I think it is a quality chair for the price assuming the above negatives aren't deal breakers for you. I will be keeping this chair since I do not spend a lot of time at my desk and the overall quality outweighs the few negatives for me.
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Reviewed in the United States on September 27, 2016
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