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Description
Human STAT4 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. |
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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 Signal Transducer and Activator of Transcription 4 (STAT4) capture antibody. After incubation and washing, the assay 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 Signal Transducer and Activator of Transcription 4 (STAT4) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Signal Transducer And Activator Of Transcription 4 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Signal transducer and activator of transcription 4 (STAT4) is a transcription factor belonging to the STAT protein family, which consists of STAT1, STAT2, STAT3, STAT5A, STAT5B, and STAT6. STAT proteins are key activators of gene transcription, binding to DNA in response to cytokine gradients. STAT proteins are common components of the Janus kinase (JAK) signaling pathway and are activated by cytokines. STAT4 is required for the development of naive CD4+ T cells into Th1 cells and for the production of interferon-γ in response to IL-12. There are two known STAT4 transcripts, STAT4α and STAT4β, which differ in their downstream interferon-γ (IFN-γ) production. | |||||||||||||||||||||||||||||||||
| 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, and other biological fluids |
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Product Reviews
★★★★★ 4
Good bag for a good price
Size: Large, Color: Amourpink
Really loved this backpack. I got it after reading all the reviews and wanting something for travel. The inside pockets with extra compartments that are water proof or organizing was a really nice touch. I traveled on airlines with this a few times and then started using it for school as well. It help up really well. I tend to be kind of rough on things and unfortunately the zipper broke. I'm unsure of if I over stuffed it or it was just a cheaper zipper. I would have considered resorting it because I used and loved it so much but my mom's dog got to it and it was not salvageable after that. I would recommend this specifically as a great travel backpack for the air port but it can easily double as an over night bag or school back pack.
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Reviewed in the United States on April 13, 2026
★★★★★ 5
So big, but lightweight!
Size: Large, Color: Amourpink
I bought this backpack to have as my "personal item" on the many flights I have to take. It is the perfect weight, having a place for just about anything you'd want to have during a flight, as well as having your carry-on items. The charging port is a bit of a challenge to plug in because of how you have to position the cord, but it is doable. I did not end up using the charging feature on my most recent flight, but I can see how it would be helpful for the travel that does not provide many charging stations. I have some serious back issues, but even full, I found this backpack to be a reasonable weight. I cannot attest to it's durability at this time, as I've only taken it on two trips now, but it is a great bag. There are many compartments, which can be good and bad. I recently put my wallet with all my identification and cards in a compartment that was easily missed. I thought I had lost my wallet and was panicked about how I'd get on my return flight if I didn't find it. After completely emptying the bag in my friend's car, and running my hands all over the compartments, I found the wallet. Here's the thing, my wallet is not little, so you can see that the walls of this backpack are pretty thick. The pink color is more of a dusty pink, which I find to be more attractive than a bright pink for something like this.
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Reviewed in the United States on May 22, 2026
★★★★★ 5
Durable and easy to carry
Size: Large, Color: Amourpink, Size: Large, Color: Amourpink
I absolutely love this backpack! I got it for our two weeks long Northern Europe trip. It is very lightweight, spacious, and surprisingly comfortable even when fully packed. The compartments are well designed and make it easy to organize everything for travel. I especially liked how convenient it was for airport trips and fitting over the carryon’s handle and under the airplane seat. The material feels durable and the overall quality exceeded my expectations for the price. Stylish, practical, and perfect for short trips or everyday use. Highly recommend!
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Reviewed in the United States on May 19, 2026
★★★★★ 5
Very good product
Model: iPad Mini 6/A17 Pro 7th Gen
Nicely attaches to the iPan. Easy and no bubbles. had one and worked it completely out. Had all the need items for the installations. Screen touch works, cover went on without a hitch.
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Reviewed in the United States on May 25, 2026
★★★★★ 5
iPad mini screen protector, good price range.
Model: iPad Mini 6/A17 Pro 7th Gen
Good coverage, easy to apply. Thorough dust removal. I have paid much higher price at retail. Good quality.
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Reviewed in the United States on May 25, 2026