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Description
Human RBBP7 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. 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 the collected cells three times with ice-cold PBS and resuspend them 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 collect the supernatant for analysis. 5. 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 a capture antibody against retinoblastoma-binding protein 7 (RBBP7). 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 retinoblastoma-binding protein 7 (RBBP7) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Retinoblastoma-binding protein 7 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Piston-binding protein RBBP7 (RBBP7) is a protein encoded by the RBBP7 gene. This protein is a ubiquitously expressed nuclear protein belonging to a highly conserved subfamily of WD-repeat proteins. It is found in several proteins that directly bind to the retinoblastoma protein, which regulates cell proliferation. The encoded protein is present in many histone deacetylase complexes, including the mSin3 co-repressor complex. It is also present in protein complexes involved in chromatin assembly. | |||||||||||||||||||||||||||||||||
| 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 homogenates, cell lysates and other biological fluids |
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4.2 ★★★★★
Based on 21 reviews
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Product Reviews
★★★★★ 5
It Works!
Size: Full-size Truck, SUV
When I bought my vehicle it didn't come with a spare. I knew what I was in for. I was on my way to an important meeting and as fate would have, the tire pressure indicator reported a flat. I imagined the worst but experienced the best!
The directions said to let all the air out of tire and remove the valve stem core, it seemed like I was making a bad situation worse. Then it said to attach the tube to the stem and squeeze the contents into the tire. I thought "this is never going to work" the fluid seemed too watery to seal anything, but I followed instructions, I reinstalled the valve core and reinflated the tire, and it worked!
I expected it to leak immediately, it didn't. It lasted all day and most of the next day until I could get the tire to the shop. They removed a nail and patched the hole, overall I'm very satisfied, I bought a replacement right away, just in case!
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Reviewed in the United States on May 20, 2026
★★★★★ 5
Better than the green goo
Size: Full-size Truck, SUV (2-pack)
Recently, while I was at work, I noticed that I had at some point run over a nail with my car. Normally, you'd take your car to the tire shop and let them plug the hole for you. Well, the only problem was that the nail decided that it was going to poke my tire where the sidewall meets the tread, and most shops will refuse to plug a tire when it's punctured there. Makes sense, it's a liability issue for them, so I can't blame them. That still leaves me with the problem of having a tire with a nail in it. So, I could either replace the tire with either a new or used tire, which is expensive or I can try using a sealant to try and get as many miles out of my tire as I can.
In the past, I had tried using the green goo sealant on nail holes, and while it works to get you 15 miles down the road, that's about it. Now, while TireJect doesn't recommend using their product on punctures on the shoulder of the tire, I decided that I'd give it a try to see if it would work. And as of writing this review, it's been about 10 days since I put TireJect in my tire, and so far, it hasn't leaked any air, despite it not being meant for the shoulder. When money isn't so tight, I'll probably go ahead and replace the tire, especially since there's only about 4/32 tread depth left, but for the time being, I'll keep running it, and hopefully it keeps holding pressure properly like it has been.
Now, as for how easy it is to use, the only tools you will need that aren't provided, is an air compressor to allow you to re-inflate the tire. The kit comes with everything else you will need, including a valve core removal tool and a syringe to add the sealant to your tire. To use this product, you will first need to carefully remove the valve core with the provided tool, careful not to lose the core, since it doesn't come with a replacement core. After that, thread the hose on the syringe on to your valve stem, and then carefully add the required amount of sealant to the syringe. If you accidentally spill some, wipe it off before it dries, so that it doesn't stain anything. From there, just push the goo into the tire with the plunger, adding more to the syringe as necessary. When you're all done, re-install the valve core into the valve stem snuggly, careful not to over-tighten, and air up the tire to the recommended PSI with your compressor. After all that is done, go take a short, slow drive to distribute the sealant, and then recheck your air pressure.
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Reviewed in the United States on July 29, 2025
★★★★★ 1
TPMS Compatability is a LIE
Size: Sedan, Crossover, Small Truck
2018 Subaru Impreza, with one tire with a slow leak (about 2 PSI a week) that I couldn't trace.
I bought the "Full-Size Sedan, Small Truck, Crossover" kit (the 6 oz. pouch) based on my tire size, 225/40/18.
After I removed the valve core, attached the dispensing hose and opened the cap, the product would not flow into the tire. I squeezed the pouch, but that only made the product leak out around the valve stem onto the wheel and driveway. I tried rotating the tire so the valve stem was at the top, and and also at the bottom. I even took the wheel off so I could get a fully vertical flow. Nothing worked. I finally gave up after only about half the product was dispensed, and I do think most of it went somewhere other than in the tire.
I reinstalled the valve core, remounted the wheel, plugged in my little lighter socket-powered compresseor, and tried to inflate the tire. And ... NOTHING. The pressure reading on the compressor immediately shot up to over 100 PSI, and it automatically shut off. Tried multiple times, with the same result. I again removed the valve core, and a) no air came out of the tire when I removed it, and b) even with no valve core the compressor still would not pump air into the tire. Clearly the gunk had completely blocked the valve/TPMS.
Fortunately, when I took the tire to Costco (where I'd bought it), they were able to fix the damage I'd done, as well as patching a small nail hole in the tread I hadn't seen (that was clearly the source of the slow leak) for no charge. Since I've already submitted a refund request, ultimately all it really cost me was my time and effort ... and a whole lot of stress.
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Reviewed in the United States on November 26, 2024
★★★★★ 5
Forget the gooey stuff, this product WORKS! See UPDATES 1 and 2
Size: Compact Car, Mid-size Sedan
I've had a slow leak in one of my tires for some time. It's a common thing with chrome plated alloy wheels, they get corrosion on the bead area where the tire seals and then you get a slow leak. This was the only product which is water soluble so it's not going to make a permanent mess inside the tire; additionally, it contains tiny bits of Tufsyn which is the material some tires are made from. So it's not just a liquid that is sealing the leak, it's tiny bits of rubber. I was putting 5 or 6 lbs of air in my tire every few days, and I could see that the tire was getting low. I installed this TireJect per the instructions a week ago and the tire appears to be doing great, despite the fact that we've had some very cold weather. (I didn't want to check it with a gauge while it was this cold out because just in case the tire's valve didn't seal all the way; I'll check with a gauge when this cold snap is past.) While I can't say for 100% sure that there is no more leak, I suspect that the leak has been sealed up completely. This is a product unlike the others and I think it's worth the price.
FIRST UPDATE: It has been two months and I haven't had to add any air at all to the tire where I used this product. There was a slow rim leak which was losing about a pound or so per day and it has been totally fixed. The car has sat for several days a few times, including in single digit cold weather, as well as driven regularly otherwise, and the tire hasn't lost any air. I think it's the micro Tufsyn fibers in this product which seal leaks better than some goopy glue stuff other brands are made of. I totally recommend this product!
SECOND UPDATE - it's been eleven months since I added this to my tire with the slow rim leak and I have not had to add air a single time. Not once. THIS STUFF WORKS. Forget the slime and foam and get this.
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Reviewed in the United States on December 5, 2024
★★★★★ 5
Good
Prefect. It’s exactly what my lawnmower needed.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 2, 2026