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Description
Rat RUNX2 ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 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 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 Runt Related Transcription Factor 2 (RUNX2) capture antibody. 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 Runt Related Transcription Factor 2 (RUNX2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Runt Related Transcription Factor 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Runt-related transcription factor 2 (RUNX2), also known as core binding factor subunit α-1 (CBF-α-1), is a protein encoded by the RUNX2 gene. RUNX2 is a key transcription factor involved in osteoblast differentiation. This protein, a member of the RUNX transcription factor family, possesses a Runt DNA-binding domain. It is essential for osteoblast differentiation and bone morphogenesis. It serves as a scaffold for nucleic acids and regulatory factors involved in skeletal gene expression. The protein can bind to DNA either as a monomer or as a subunit of a heterodimeric complex, with a stronger affinity. Transcript variants encoding different protein isoforms result from the use of alternative promoters and alternative splicing. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids |
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4.8 ★★★★★
Based on 21 reviews
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Product Reviews
★★★★★ 5
Works
Size: 3 Panel-102'', Color: Beige, Size: 3 Panel-102'', Color: Beige
It’s beige and not white. Once install - hard to disinstall. Need a drill to put it together
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Reviewed in the United States on May 4, 2026
★★★★★ 3
Does the job, but assembling by yourself is a nightmare
Size: 4 Panel-88'', Color: Black
Does it do the job? Yes, although as others said there are small gaps but it's not a huge deal. The price is also good. But the reason I'm giving it a 3/5 is simply because the assembly for this was a complete nightmare. I honestly don't think I would recommend this to anyone unless they have another person to help them assemble it, because doing it by myself was terrible. I don't think I'd buy this again, I think I'd opt to just spend a bit more money and save myself the trouble personally.
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Reviewed in the United States on May 29, 2026
★★★★★ 4
Reasonably adequate room divider
Size: 4 Panel-88'', Color: Beige
I'm reviewing this as I assemble it.
Couple things:
1. I didn't expect as much assembly. I've ordered dividers before and they more-or-less came as one unit. Sometimes the panels needed screwing together. These require complete assembly and come largely as three rods: two make up vertical columns and snap together. Another one (called part "C") makes the horizontal columns and you have two of these per panel (one attaches to part "A" and the other part "B"). These parts are metal with a plastic shim. Using the wood screws to attach to part "C" is a real pain in the neck. There's not much holding the panel in place so it's a little tricky.
One tactic I've found while I'm assembling that works for the initial connections from parts A and B to their respective "C" rods is to hold the screw in place with a screw driver and then rotating the rod around the screw. This will do a number on your hands if you aren't wearing gloves. This obviously doesn't work when completing the connection.
Using a driller driver on this is really near impossible because there isn't anything you can use to secure it in place. You can use it on the first panel, but as it gets longer, it becomes increasingly difficult and because it isn't wood, it's really tight. I considered drilling larger pilot holes but since there are only 4x4=16 screws I need to screw in, I just decided to use my screw driver to complete it.
2. Also related to assembly. When completing the panels (attaching parts "A" and "B" to parts "C" that have the cloth cover on it), you have to be careful that when you tighten that side that it isn't loosening the other side. Because the pilot holes are so tight, you can end up rotating the rod, which rotates it in the same direction as looser on the original side. Having someone hold the "C" rod in place while you screw it in is probably the easiest approach. I didn't have a 2nd person, so I just had to keep flipping back and forth and tightening both sides as I screwed it in. Not the worlds biggest deal, but annoying nonetheless.
3. The way the instructions are written, they seem to suggest building this thing progressively; that is, you do panel 1, then 2, connect them together, then do 3 and connect it, etc. I took a different route that I suspect saved me quite a bit of trouble, and I assembled all four panels first and THEN connected everything together.
4. For the love of God make sure you check that the plastic tip is on the same side for every panel. Otherwise, you have to take one side apart again and reverse it. On the bright side, if this happens, you've essentially bored out the pilot holes to be the correct size... which is having me question if I shouldn't have just bored them out to the appropriate width in the first place.
5. Attaching all of the panels together is also an enormous pain in the ass unless you happen to have an 88" long elevated surface. Attaching the legs either requires you to elevate one side, which will invariably twist the inexplicably cheap material in the bottom connectors... or you can attach them sideways... or you can put this thing upright, having two people hold the panels in place while you use the allen wrench to tighten the bolts on the underside. None of those are particularly great options.
NOW on to the utility itself.
1. The panels do let some light through (I didn't believe their advertising, and that was one of the reasons that I bought beige, is that I wanted it to not be too dark). They aren't transparent though, so it isn't that far off from their description. They functionally work great, and keep the mess of wires hidden and when I'm sitting at my desk, actually reflect quite a bit of light into my office. Great!
2. My wife has described these as "the most hideous piece of furniture ever conceived of by man." So it does not have spouse approval factor. Granted, she will seldom be in my office area, so that isn't the end of the world.
3. These are really hard to align in a way that doesn't look a little tacky. There are some plastic connectors but they don't do a bang up job of keeping these in place. Each panel is slightly tilted and it's... quite obvious. I may at some point make my own improvements to these to help make them more level. It's not a particularly expensive product so I wasn't expecting much so it's fine and I'm not going to ding them on the rating because of it.
All said, would I buy this product again? Probably not. It's assembly was ~90 minutes which is about 75 minutes longer than I was anticipating spending on this (not including the 5 minute writeup that I'm doing here). But am I going to return it? Also no, if for no other reason I'd be just as annoyed taking it apart and putting it in the original box to return it.
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Reviewed in the United States on October 31, 2023
★★★★★ 5
Great product
Color: Black, Size: 4 Panel-88''
Literally exactly what I was needing
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Reviewed in the United States on May 1, 2026
★★★★★ 5
It's good...
Color: Black, Size: 4 Panel-88'', Color: Black, Size: 4 Panel-88''
It's good....took about 45 minutes to put together. No issues. Looks great...made the wife happy....
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Reviewed in the United States on May 14, 2026