Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Jul 8 - Jul 13
Promo Codes Available:
For Your Every Summer RSVP, with Code: SUMMER15
Description
Human NCOA3 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: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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. 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. |
|||||||||||||||||||||||||||||||||
| 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 Nuclear Receptor Coactivator 3 (NCOA3) 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 Nuclear Receptor Coactivator 3 (NCOA3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Nuclear Receptor Coactivator 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
|
|||||||||||||||||||||||||||||||||
| Background | Nuclear receptor coactivator 3, also known as NCOA3, is a protein encoded by the NCOA3 gene. It is also commonly known as "amplified in breast 1" (AIB1), steroid receptor coactivator 3, or thyroid hormone receptor activator molecule 1 (TRAM-1). It is a transcriptional coactivator protein that contains several nuclear receptor interaction domains and intrinsic histone acetyltransferase activity. | |||||||||||||||||||||||||||||||||
| 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. |
|||||||||||||||||||||||||||||||||
| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
Shipping Notes
- Free Standard Shipping on $100+ Orders to the USA.
- Except Preorder products are shipped in 48 hours.
- Delivery to the USA:
- Standard Shipping : 3-10 business days
- If time is of the essence, please consider selecting expedited delivery for faster service.
Exchange/Return Notes
- We offer a 30-day return/exchange service after receiving.
- Final sale items are not eligible for returns or exchanges.
- To process your return/exchange, please contact us at [email protected]
- Please click here for more details>>> Return & Exchange Policy
4.1 ★★★★★
Based on 1209 reviews
Sort
Product Reviews
★★★★★ 5
My every day watch. I love it.
Color: Silver/Black/Gold
Is it the nicest watch? Nope.
Does the clasp come undone as time goes on? Yep.
Do I still wear it every day? Absolutely.
This is my daily driver watch. It will win no awards in quality or style, but it looks great, goes with everything, has minimal error as days go by, and is easy to read. Cheap and great. A rare combo these days.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 31, 2026
★★★★★ 5
Great value for dress watch or casual use
Color: Silver/Black/Gold, Color: Silver/Black/Gold
Amazing piece for the price. Had many compliments from coworkers and family about the watch, a real conversation starter. second hand is on point and even with big wrist it suits me well
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 23, 2026
★★★★★ 5
Great quartz Datejust alternative!
Color: Silver/Black/Gold, Color: Silver/Black/Gold
I was looking for a cheap quartz Datejust alternative and many online reviews suggested this watch. I originally wanted the black and silver version but as prime only had the smoky gray and Gilt indices I went with that, in case it didn't meet expectations and needed an easy return. I needn't have worried. This watch is great!
The smoky gray sunburst dial is beautiful (as good as my Aqua Terra), and matches well with the Gilt applied indices and hands and white printing (words and minute ring on border); the Gilt details give it the vintage heritage look that's popular now. It's also very easy to read (as opposed to my silver Omega quartz) and date only (which oddly makes it seem pricier). Lume is fair and present on the hands alone, but better than my Tag Heuer link. The folded bracelet is a little stiff but easy to size (google how to size a folded bracelet and this was about the easiest I've done), good quality and very comfortable. it also has 4 microadjust positions on the clasp. The movenent also hacks.
On the wrist, it looks very similar to the datejust but without the heft and probably better sized at 39mm (with 20mm band). The smaller sizing and price also make it a nice alternative to the Tissot gentleman quartz. And the Casio brand makes it probably one of the cooler datejust homages, as its the brand that both hipsters and billionaires wear (per style and value).
Downsides are the plated case (not full stainless), folded bracelet (though to be fair Rolex used folded bracelets originally as well) and snap on back, but at $30 can you really complain? And I needn't worry if I accidentally bash it against a doorsill. I'm wearing it already!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 8, 2020
★★★★★ 4
Great quality/ customer support
Color: Silver/Black/Gold
I had to order a replacement because the first box was damaged and had been previously opened. Thankfully it’s was super easy and fast to do, i got the new one and is amazing, its chic and classic. It’s the perfect size for small writs!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 18, 2026
★★★★★ 5
Serves the purpose
Color: Silver/Black/Gold
Looks very nice for an inexpensive watch. And keeps time.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 1, 2026
recommand products
RockShox Super Deluxe Select+ Rear Shock 230x65mm
157.00
【新品】POP|CULTURE 5 / Alstroemeria Records 発売日:2015年12月28日
24.00
DT Swiss M 1900 Spline Aluminum Tubeless 27.5" Front Wheel
169.99
RockShox ZEB Ultimate 29" 180mm Fork 15x110mm 44mm Rake
350.00
DT Swiss ARC 1400 DiCut Disc Carbon Tubeless 700c Wheelset
700.00