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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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4.5 ★★★★★
Based on 1280 reviews
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Product Reviews
★★★★★ 5
Transformative!
Format: Paperback
This book is well written and extremely informative! It has definitely helped me to take the energy work I do to a whole new level I actually recommend all the books in this series!
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Reviewed in the United States on May 8, 2026
★★★★★ 5
A Healer's Book
Format: Paperback
In my opinion, this book is most suited to the "healers". I am only part-way through it right now, but thoroughly agree with what I have read so far because I have either experienced it or it makes logical sense from where I am on my personal journey. The book is slow reading, not difficult, but slow if you really want to understand what Barbara has written as she combines science with metaphysical beliefs. I cannot compare this book to her first book because, although I purchased her first book years ago, I honestly have not read it yet. "Light Emerging" was recommended by my master teacher. Barbara has spent significant time studying the auric fields of people vs. behavior.
I plan to return to this review at a later time and update it with additional information, but I see it as a valuable resource for a healer (for both healing self and healing others). By the way, if you don't know it already, healing is not limited to the physical body. There is emotional healing and spiritual healing as well. ALL are important. The healthiest people are those who are balanced on all levels.
This 342-page book explains a lot about energy, both scientifically and metaphysically. If you have already done significant reading, this book expands on a lot of it, such as explaining the chakras and the movement of energy throughout the system. As Barbara goes through each chakra, she explains how you likely behave if that chakra is fully functioning or if it is blocked. Quite a few illustrations within the book visually show you the energy movements within people who exhibit certain types of behavior, and the movements of energy within groups of 2 people exchanging energies at certain levels. This is one way in which this book is different from many other chakra and energy vibrational books. An example is an illustration of a couple in dispute and how they energetically resolved the dispute. Barbara has the developed ability to see the auric fields, as some do, and she supplements her writing with experiences she has had with clients. In this instance, she explains the energy movement in 3 stages of the therapy between the couple, as well as illustrates it in picture form, so you can follow what is happening energetically between the couple throughout the process.
Here is a little tid-bit for you from the book: Barbara mentions that Dr. Robert Beck, a nuclear physicist, found that all healers exhibit the brain-wave patterns of 7.8 - 8 Hz while they are giving healings, the same fluctuation of the earth's magnetic energy field (the Schumann waves). The assumption is that the healers are synching up with the earth's magnetic field when doing healing. Dr. John Zimmerman, founder and president of Bio-Electro-Magnetics Institute, found through additional research, that once healers have linked with the Schumann waves, the right and left hemispheres of their brain have become balanced with each other and show a 7.8 - 8 Hz alpha rhythm. After they link with the patient for some time through laying on of hands, they are able to link the client in with the same earth's energy magnetic pulsations.
Good book. If you are into metaphysical energy and healing, I think you will enjoy it and find it very informative. If you are NOT into metaphysical energy and healing, you will likely not understand it and may get nothing from it, especially if you are not open to this area of study.
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Reviewed in the United States on August 31, 2008
★★★★★ 5
Sequel to Hands of Light
Format: Paperback
Hands of Light has been widely recognized as the foundational text merging the Eastern approach to healing through cleansing and balancing a person's chakras and auras with the Western approach to healing through re-tuning a person's electromagnetic frequencies by applying the principles of quantum physics.
Barbara's sequel, Light Emerging, further amplifies and extends these concepts. Specifically, she elaborates on the applications of the Holographic Model, the Creative Process and the mechanisms of Spiritual Guidance. She defines and diagrams the four dimensions of humanity at the Physical, Auric, Harac and Core Star levels. Through these four dimensions, average humans, once healed and balanced, can mature to be channels of energy, connecting heaven to earth vertically, and connecting all of humanity horizontally. The whole of Barbara's teaching can be incorporated into these four simple, clear and deeply profound diagrams.
Since her previous book, Barbara set aside her private practice to establish a four-year university-level program to train and accredit healing practitioners to apply these disciplines. This book incorporates the further insights and experiences she gained through developing her School. Light Emerging is written specifically to instruct readers on how to self-heal and to give readers a preview on the commitment required to becoming a healing practitioner. This book is not intended to replace her School's four-year degree in healing. Appendix C offers links to current lists of her practicing graduates, by city and state.
There is no duplication between her books. I personally benefited by reading and studying them side by side. I have also called a number of her graduates here in California and confirm they have both the meticulous training and proper motives to follow in Barbara Ann Brennon's footsteps.
No fiction here. This book exemplifies the theory and practice of genuine physical, psychological and spiritual healing, either through self-study or through a practitioner.
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Reviewed in the United States on August 24, 2011
★★★★★ 5
only you can initiate the healing response
Format: Kindle
We are all divine light and this book gives you the tools to access your own healing response to live your life in light and love.
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Reviewed in the United States on November 5, 2025
★★★★★ 5
Good deal
Format: Paperback
This book is in fine shape. Like new
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Reviewed in the United States on May 10, 2026