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Description
Human TBX21 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 handling and requirements: Cell lysis buffer: Gently wash adherent cells with ice cold PBS, then trypsinize and harvest by centrifugation at 1000g for 5 minutes. Suspension cells can be harvested directly by centrifugation.
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 handling and requirements: Cell lysis buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and harvest by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Wash harvested cells three times with ice-cold PBS and resuspend in 150-200uL 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. The extract was centrifuged at 1500×g for 10 min at 2-8°C, and the supernatant was collected for detection. 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 the T-box transcription factor TBX21 (TBX21). After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of T-box transcription factor TBX21 (TBX21) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human T-box transcription factor TBX21 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | T-box protein 21 (TBX21), also known as the T cell-specific T-box transcription factor T-bet (TBET), is a protein encoded by the TBX21 gene. This gene is a member of a phylogenetically conserved gene family that shares a common DNA-binding domain, the T-box. T-box genes encode transcription factors involved in the regulation of developmental processes. TBX21 is a Th1 cell-specific transcription factor that controls the expression of the hallmark Th1 cytokine interferon gamma (IFNG). It regulates the development of naive T lymphocytes. | |||||||||||||||||||||||||||||||||
| 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 | Cell lysates and other biological fluids |
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4.3 ★★★★★
Based on 837 reviews
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Product Reviews
★★★★★ 5
Labrador Retriever Fun Toy
Color: Yellow
Excellent for older Labrador Retriever with a soft mouth. 2 levels of movements. Colorful. Recharge quickly too, cable included. 2 velcro type covers.
Fairly chew resistant, watch your dog, as with any toy! Good size, no worries about my Labrador swallowing it!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 23, 2026
★★★★★ 5
Great engaging toy for my border collie
Color: Blue
I have an 8 yr old border collie with an injured rear hock, so she can't chase the Frisbee any longer. This toy, when set on high, kicks her hunting instincts in, and she chases it, brings it back, and keeps it between her paws until it activates *or she bumps it). The plastic 'holey' covering let's her get a good grip on it, and it is a little larger than a baseball. Good quality plastic/soft rubber covering.
It charges quickly, and has unpredictable jumps and rolls unlike some others that just run on 1 track. She would play with it all day if I let her. She brings it to me to play with her, but it doesn't really need a human's intervention. A tooth will occasionally hit the small control and turn it off, but no concerns about it falling apart or her ripping it apart. Highly recomment
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Reviewed in the United States on February 26, 2026
★★★★★ 3
Not as durable as it could be, but fun
Color: Yellow
Our 55 lb lab has fun with this toy, but it's not lasting long. The ball itself is hard plastic and the yellow rubber wrap around it has already been chewed through n several places. It did come with a fabric cover, but that, too, is not durable. Would not recommend leaving with your dog unattended as they could eat the rubber off in no time. But to bring out occasionally while watching the dog, which is what we do, it does provide some fun for all!
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Reviewed in the United States on May 17, 2026
★★★★★ 4
Durable enough for my dog's tough play. She loves this interactive ball.
Color: Blue, Color: Blue
I love these interactive ball toys. My dog gets so excited and engaged with them. I went through 3 of them that my dog destroyed in a short time. They simply were not durable enough for her tough play. So far, we have had this one for a few weeks, it is holding up and withstands her chewing and rough play, and she loves it. It keeps her busy for a long time.
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Reviewed in the United States on May 7, 2026
★★★★★ 5
Great toy for old dogs
Color: Yellow
Great to bring a old dog back to his old self. Have a 10 yr old who has a little shoulder pain but needs a little light excitement. This did the trick, he paws it a little and enjoys moving it around. It can take his biteing without breaking.
Hopeing our small dog whos afraid of it will watch and get over her fear. She learns everything from him.
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Reviewed in the United States on May 6, 2026
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