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Description
Human GR-a 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 Glucocorticoid Receptor Alpha (GR-a) capture antibody. 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 Glucocorticoid Receptor Alpha (GR-a) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Glucocorticoid Receptor Alpha ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Glucocorticoid receptor α (GR-α) is one subtype of the glucocorticoid receptor, the other being GRβ. Both are derived from the same transcript of the glucocorticoid receptor gene, spliced differently. GR-β can bind to DNA but not to steroids, potentially acting as a steroid inhibitor by interfering with GRα's DNA binding. GRα and GRβ are identical in their first 727 amino acids. From amino acid 728 onward, GRα has a 50-amino acid sequence, while GRβ has only 15. At the mRNA level, both contain exons 1 through 8, differing in that GRα contains exons 9α and GRβ contains exons 9β. GRα is expressed in virtually all tissues and cells, and its level far exceeds that of GRβ in most cells. Glucocorticoids primarily exert their biological effects by binding to GRα. | |||||||||||||||||||||||||||||||||
| 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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4.8 ★★★★★
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Product Reviews
★★★★★ 4
Content is better than the title!
Format: Paperback
The best quality of Heiser as a scholar/writer in my opinion is that he appreciates the academy but does not acquiesce to its influence. This is seen most in his correct interpretation of the identity of the sons of God in Gen 6 and the identity of Satan in Is 14 and Ez 28 etc. which were long abandoned and scorned by the academy.
All 60 articles are well written with clarity and brevity!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 6, 2019
★★★★★ 5
Great Read
Format: Paperback
An outstanding collection of thought provoking essays. This makes a great daily 5-10 min read.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 14, 2026
★★★★★ 5
Read all of his books, you won't be dissapointed.
Format: Paperback
I love Dr. Heiser books, but I wanted to step back from the spiritual realm and reconnect with scripture again. This book is light, easy to read, and like all his content, so eye opening. I am alway slapping myself in the forehead and wondering how I didn't know or notice things. "I Dare You Not to Bore Me With the Bible" is another of his books that I'm reading simultaneously. If you want to go deeper in Bible Study, this is the book. It's helped me forgive a lot of the Bible "hangups" I have that were very cultural or significant for reasons I missed. Get those 2 books (this and the one I mention above), and then when you're ready to really have your understanding of G-d and the Bible blown out of the water, move on to Unseen Realm, Reversing Hermon, Supernatural, and all of the others. If you desire to understand the Bible, and not just show up on Sunday, read everything by Dr. Michael Heiser, his website is a great resource too. So grateful for the work he does.
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Reviewed in the United States on May 6, 2021
★★★★★ 5
Great Read
Format: Paperback
Just started reading it, but seems like it’s going to be very good.
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Reviewed in the United States on October 14, 2025
★★★★★ 5
Excellent tool for gospel workers!
Format: Kindle
For those of us who want to contextualize the gospel in pagan contexts, Dr. Michael S. Heiser helps us get beyond our own cultural context to the ancient cultural context of the Bible itself. One could spend years and years researching the ancient contexts of the Bible, but for workers with families and careers other than Dr. Heiser's, the time needed would far exeed what is practical.
Thankfully, there is a solution: Dr. Heiser has done a lot of the distilling needed to grasp the basic cultural and linguistic backgrounds of the Bible writers. After presenting the Unseen Realm, Reversing Hermon and I Dare You Not To Bore Me With The Bible, he has done one better here in the Bible Unfiltered. Why do I say better? Well, I would say that as time has gone on, Dr. Heiser's ability to communicate complex topics for the average layman has improved markedly.
Get this book, let it whet your appetite, and then please do get Unseen Realm and Reversing Hermon. They will help you to understand the Bible, contextualize the gospel, and offer an entrance to a fantastic intellectual and spiritual journey of a lifetime.
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Reviewed in the United States on January 3, 2018
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