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Description
Human FOXA2 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 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 Forkhead Box Protein A2 (FOXA2). 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 Forkhead Box Protein A2 (FOXA2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Forkhead Box Protein A2 | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Forkhead box protein A2 (FOXA2), also known as hepatocyte nuclear factor 3-β (HNF-3B), is a transcription factor that plays important roles in development and mature tissues. When dysregulated or mutated, it also plays a crucial role in cancer. FOXA2 belongs to a subfamily of forkhead box (FOX) transcription factors, the other members of which are FOXA1 and FOXA3. These proteins were originally named hepatocyte nuclear factor 3α, β, and γ. These transcription factors contain a forkhead domain (also known as a winged-helix domain) flanked by sequences essential for nuclear localization. Their N- and C-termini are also conserved, serving as transcriptional activation domains. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.7 ★★★★★
Based on 14 reviews
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Product Reviews
★★★★★ 5
Very good speakers
Size: Pair
The r51ms are used to compliment my klipsch rp500m speakers and a 12 inch subwoofer in a large living room. By placing the r51m speakers to the outside walls , and the rp500m speakers to the interior, closer to the amplifier, and with the subwoofer set on deep bass only at just enough volume to barely compliment the sound, the large living room is more than sufficiently covered with at least true high fi quality sound, if not true budget audiophile sound. My entire 2.1 system is all klipsch. That's how I want it.
I am using them with a high powered tube amplifier, so presence and clarity is awesome. Placement is critical. I was initially disappointed until I switched the r51ms to the spread to the outside walls position, and the rp500ms to the interior, closer to the amplifier. The difference was remarkable. The 2 different klipsch speakers, ie one a rp and the other a r series blend together perfectly with no clash whatsoever because of somewhat more refined sound on the rp500m speakers. The rp500m speakers do sound higher end , but they are supposed to, but with the speakers as outer r51m and inner rp500m, and the klipsch subwoofer at deep bass with just a touch of volume the result is sound of such quality that I can honestly say I can be content and satisfied with it. I can say my system is complete, and no further improvement, upgrades are needed. Tower speakers as primaries are not necessary because of the high quality of sound and the high sensitivity of the 4 klipsch bookshelf speakers and klipsch subwoofer. This saves me money during this rapid inflation period of time. I could easily say that what I have sounds more than good enough to be satisfied and content with without any desire to upgrade further at this point.
Shipping speed was excellent via Amazon itself, and the box and speakers arrived in handled with care and new condition.
I am not disappointed at all now that the r51m speakers are the outside speakers, spread apart from corner wall to wall to compliment the interior placements of my rp500m speakers by klipsch. Im all klipsch here and I love it. 5 stars categorically. As previously stated, I see no need for primary tower speakers, even in a large living room, and that equals high quality sound and savings of money at the same time. I recommend these speakers. They do more than just do sound quality wise.
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Reviewed in the United States on August 22, 2025
★★★★★ 5
Awesome speakers.
Size: Pair
Love Klipsh speakers. Beautiful, full, rich, clear sound. Combine with a sub for best sound. Very loud. Love these.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 30, 2026
★★★★★ 5
Love it
Size: Pair
These are nice for any audiophile. They look and sound amazing.
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Reviewed in the United States on May 14, 2026
★★★★★ 5
Great Sound
Size: Pair
Great things sometimes come in little packages. Powerhouse sound with virtually no input at all. Tight, crisp, and well-made as usual from Klipsch. If you are looking for a powerhouse monitor these are what you want.
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Reviewed in the United States on April 22, 2026
★★★★★ 4
Great starter speakers - for the right price.
Size: Pair
A good all-around pair of starter speakers if you're planning on using them as standalones. These will have zero trouble filling a small to medium sized space with rich and detailed sound at low to mid volumes. The bass is adequate for their size. They're not going to knock your socks off on more thumpy genres. I would imagine pairing them with a subwoofer would make quite the difference but for rock and some synth pop, they're pretty sweet. Plus the copper woofers look very striking without the faceplates.
That said, I say these are best suited at low to mid volumes because cranked, the higher frequencies can be awfully fatiguing coming out of these speakers. I would not pay anywhere close to the $349 MSRP for these (you can probably find a pair of far superior RP-600Ms for around that price nowadays). Around $150 seems like a sweet spot for the R-51Ms, but the Sony SSCS5s and Fluance Signature bookshelfs are very comparable, if not better sounding for around the same price. Still, these will probably more than satisfactory if you're not expecting to be blown away by them.
UPDATE 5/20/25:
Can confirm that the subwoofer made a big difference. I was going to retire these since I replaced them with some bigger floor speakers on my main setup but I decided to try them out in place of the factory speakers on the Klipsch ProMedia 2.1 I have hooked up to my computer. Not only does the onboard amp seem to drive them okay at not-insane levels of volume - they sound really sweet paired with the 6.5-inch subwoofer. While I won't change my rating, I do highly recommend getting a sub along with these speakers.
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Reviewed in the United States on April 11, 2025
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