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Description
Human SP7 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 enzyme conjugate are sequentially added to microwells pre-coated with a Transcription factor Sp7 (SP7) 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 Transcription factor Sp7 (SP7) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Transcription factor Sp7 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Osterix (OSX), also known as the transcription factor Sp7 or zinc finger protein Osterix, is a protein encoded by the Sp7 gene. It is a member of the Sp family of zinc finger transcription factors and is highly conserved among osteogenic vertebrates. Together with Runx2 and Dlx5, it plays an important role in promoting the differentiation of mesenchymal precursor cells into osteoblasts and ultimately osteoblasts. It also plays a regulatory role by inhibiting chondrocyte differentiation, maintaining the balance between mesenchymal precursor cells and the differentiation of ossified bone or cartilage. Mutations in this gene are associated with a variety of dysfunctional bone phenotypes. The most direct example of its role in disease is recessive osteogenesis imperfecta (OI), a type I collagen-related disorder that causes a heterogeneous range of bone-related symptoms. | |||||||||||||||||||||||||||||||||
| 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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4.1 ★★★★★
Based on 1624 reviews
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Product Reviews
★★★★★ 5
Great product
Color: Green
It was fun to watch my dog, get the food out of the rolls of fabric
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 31, 2026
★★★★★ 5
Great puzzle, even better complimentary squeaky carrot
Color: Red, Color: Red
Our dog is a Jack Russell mix and needs near constant interaction. In the winter, it gets so cold that we can't always get him out for long enough periods of time to satisfy his poor bored little brain.
I've tried so many puzzles. He gets frustrated and bored quickly, and ends up just barking at them.
One game that he loves in winter though, is what we call "the blanket game" and it's when we hide one of his favorite toys in an old blanket, wrap it all up, and then let him loose to find the toy. He can play this game over and over, for over an hour if we have the energy. So, when I saw this ball, I knew I had to try it.
He loves it. He begs me to let him have it at least once a day and it gives him a good 5-10 minutes of play. He'd play it more if I let him have that many treats, haha.
But I had to say that, true to form, his REAL favorite part of the puzzle ball was the complimentary carrot toy that came with it.
He is obsessed with the carrot.
He carries it everywhere. It goes to bed with him. He never loses it, always knows exactly where Carrot is. He even knows the word "carrot" now, after only a week or so of having our dear, beloved Carrot.
I can ask him, "go get Carrot," and he brings it right to me.
It is the only toy he ever brings me now.
The squeaker is long dead, he killed it within the first couple days, but he still chews it relentlessly, listening for the little air puff sounds it makes now.
So anyway, the ball is great. Love it. But the carrot? Is an epic find. I included a picture so you can see how utterly filthy it is from how much he loves it (don't worry, it's going in the washer).
Thank you for this random carrot. It might be the best thing to happen to our weird little family this winter.
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Reviewed in the United States on February 6, 2024
★★★★★ 3
Basic Starter Puzzle, But Not Very Durab
Color: Red
This toy is more like a level 1 puzzle—it doesn’t take long at all for my 1-year-old, 5 lb Papillon to figure it out. The ball part is sturdy enough, but the cloth sections are very soft and not durable. I worry about him tearing into it and possibly swallowing threads, so I only let him play with it under close supervision.
If you just need a simple puzzle to introduce your dog to enrichment toys, this could work. But if you’re looking for something that lasts longer or provides more of a challenge, this isn’t it.
Pros:
• Works as a very basic starter puzzle
• Ball portion is fairly sturdy
• Good for quick enrichment sessions
Cons:
• Cloth material is soft and easy to damage
• Requires close supervision to prevent chewing or tearing
• Not challenging for dogs beyond beginner level
Overall, I’m giving it 3 stars. It does the job as an entry-level puzzle, but it’s not durable and doesn’t offer much mental stimulation.
UPDATE: The company reached out after my review and provided good customer service, which I really appreciated.
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Reviewed in the United States on September 3, 2025
★★★★★ 5
He loves it, what else is there to say!
Color: Green
My dog loves this ball. Originally we used it as it is designed and put treats inside it to knock out and keep him busy, now he just loves to throw it around and fling it by the pieces of fabric that come out. He is generally a toy destroyer so the fact that it is still arounds shows that it is strong and was for sure worth the buy. Would 100% recommend for any dog that likes to keep busy!
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Reviewed in the United States on May 15, 2026
★★★★★ 5
Pupper shakes with excitement when I take this out
Color: Green
This is a very good toy for a super busy pup but take it away after they find the treats or it's getting destroyed (the fleece) totally worth the money and the five minute prep time for a play session
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 5, 2026
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