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Description
Human AR 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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| Sensitivity | 0.18 ng/mL | |||||||||||||||||||||||||||||||||
| 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 an Androgen Receptor (AR) 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 Androgen Receptor (AR) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Androgen Receptor ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The androgen receptor (AR), also known as NR3C4 (nuclear receptor subfamily 3, group C, member 4), is a nuclear receptor that is activated by binding to any androgen, including testosterone and dihydrotestosterone, in the cytoplasm and subsequently translocating into the cell nucleus. The AR is most closely related to the progesterone receptor, which can be blocked by progesterone at higher doses. The AR's primary function is as a DNA-binding transcription factor, regulating gene expression. However, the AR also has other functions. Androgen-regulated genes are crucial for the development and maintenance of the male sexual phenotype. In some cell types, testosterone interacts directly with the AR, while in others, testosterone is converted by 5-alpha reductase to dihydrotestosterone, a more potent agonist for AR activation. Testosterone appears to be the primary AR-activating hormone in the Wolffian ducts, while dihydrotestosterone is the predominant androgen in the urogenital sinus, urogenital tubercle, and hair follicles. Therefore, testosterone is primarily responsible for the development of primary male sexual characteristics, while dihydrotestosterone is responsible for secondary male sexual characteristics. | |||||||||||||||||||||||||||||||||
| 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 ★★★★★
Based on 30 reviews
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Product Reviews
★★★★★ 4
A down bad MMC that doesn't give up. LOVED IT!
Format: Kindle
I love a down bad MMC and Lucky Beneventi was it and more!!!! I loved the love he had for Lexi and how willing he was to fight for them and give their relationship a try. But I could also understand why Lexi didn’t want to risk it. She knew that her cystic fibrosis diagnosis gave her no guarantee of how many years she'd live and she didn’t want to hurt Lucky. But seeing Lucky not giving up and showing her how important and worthy their love was, no matter the time left? It was beautiful!!!!! And that ending and bonus epilogue? My heart!!!!!! I loved it so much!!
I do wish we could’ve had more time with them dating and being happy because I did feel we spent more time with them fighting their attraction and then Lexi fighting what they felt for each other.
Also, there were A LOT of characters so it got a bit confusing trying to understand who was related to whom (so I guess at some point I will eventually go back and read the other series). However, keeping my focus on Lucky and Lexi helped me enjoy the story more.
Sweet Temptation is a fantastic beginning to the Love and Legacy series. It was definitely more heart wrenching than I was expecting but I adored the love story, the main characters' connection and how they got their HEA.
Tropes:
-Good girl x bad boy vibes
-Football player
-Baker
-Childhood friends to frenemies to lovers
-FMC has a chronic illness
-It’s always been you
-Down bad MMC
-No third act break up
-Brother’s best friend
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 6, 2026
★★★★★ 5
This book was great!
Format: Kindle
This book was absolutely amazing! I loved the relationship between Lexie and Lucky. I loved how Lucky was soo sweet with Lexie and how he knew he loved her from when they were kids. I felt horrible Lucky thought he caused her to be sick when they were kids and he lived with that guilt his whole life. The writing in this book like all of her books was great. Im addicted to this world and this crazy large family. I cant wait to read the next book in this series!
The duet audiobook narration was fantastic! The narrators did a wonderful job bringing this book and their characters to life! Tor Thom and Andi Arndt were incredible! I would highly recommend this book and audiobook.
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Reviewed in the United States on May 20, 2026
★★★★★ 3
Chronic illness rep
Format: Kindle
One of the things I love about Bella Matthews is that they are short and spicy, but long enough to develop the characters and have a overall plot with some subplots...that are little breadcrumbs for future books. However, this book didnt seem long enough to convey the enormity of cystic fibrosis. The pacing bounced around and I just didnt have the opportunity to really connect with the characters. I have read EVERY book by this author and LOVE how all of her books are interconnected. The family tree at the beginning is helpful to keep track of who is related to who.
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Reviewed in the United States on January 4, 2026
★★★★★ 5
Lucky and Lexie Will Break and Heal Your Heart
Format: Kindle
Oh my heart… Lucky and Lexie’s story was everything I hoped for and more.
Sweet Temptation, the first book in Bella Matthews’ new Love & Legacy series, is a heartfelt and emotional return to Kroydon Hills. From the very first chapter, I was pulled back into a world filled with witty banter, swoony moments, and characters that feel real and memorable.
Lexie Sinclair has spent the last year in Paris pursuing her culinary dreams. Living with Cystic Fibrosis, she has accepted that love isn’t something she can allow herself. She doesn’t want anyone to experience the pain of losing her, so she keeps her heart guarded. Her determination to protect herself and others broke my heart more than once.
Lucky Beneventi is the star quarterback, a known flirt, and her brother’s best friend. Lexie has always been his sweetest temptation, and now that they’re adults and living under the same roof, their connection is impossible to ignore.
Their chemistry has been building since childhood, and when they finally give in, it’s intense and emotional. Bella Matthews blends humor, tenderness, and heat beautifully, making every moment between them feel earned. Lucky’s quiet protectiveness and devotion made me fall in love with his character. He stands by Lexie even when she tries to push him away.
Lexie’s journey is raw, honest, and deeply emotional. Her strength and vulnerability are written with care, and there were several moments that genuinely moved me. Bella Matthews also delivers wonderful family dynamics. Seeing familiar Kroydon Hills characters as parents added a layer of nostalgia that fans will love.
Sweet Temptation is the perfect mix of heart, humor, and heat. It’s emotional, romantic, and hard to put down. This story made me laugh, swoon, and tear up more than once. Bella Matthews has delivered a beautiful romance that lingers long after the final page.
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Reviewed in the United States on October 5, 2025
★★★★★ 4
"It's a promise wrapped in a threat, and she doesn’t know it" ARC
Format: Kindle
I can always count on Bella Matthew’s books to have found family, a strong FMC, delicious spice and to make me laugh & cry in the same chapter. Sweet Temptation is no exception!
Lexie & Lucky have grown up together. Lucky plays pro-football & lives with Lexie’s brother Lincoln. Lexie is back in town, needs a place to stay, insert our favorite forced proximity trope!
Lexie doesn't want a relationship, she firmly believes that's selfish because she has cystic fibrosis and doesn’t want to cause her loved ones pain someday. She’s also the first and only girl Lucky hasn’t tried to chase… There’s wonderful banter & tension and then finally we get “one night lucky.. It's a promise wrapped in a threat, and she doesn’t know it”
I loved Lexie’s mom & Lucky’s sister for standing up for their relationship & dropping the truth bombs those around them needed to hear. Lucky’s mom for telling him, grand gestures don’t mean sh*t, when you take away her right to choose what she wants. If you love banter, group texts, spice, football romance, forced proximity -- definitely recommend this book!
Can't wait to read more of this second gen.
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Reviewed in the United States on September 24, 2025