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Description
Rat 11βHSD1 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 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 utilizes 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 11-Beta-Hydroxysteroid Dehydrogenase Type 1 (11βHSD1) 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 amount of 11-Beta-Hydroxysteroid Dehydrogenase Type 1 (11βHSD1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat 11-Beta-Hydroxysteroid Dehydrogenase Type 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | 11β-Hydroxysteroid dehydrogenase type 1 (11βHSD1), also known as cortisone reductase, is an NADPH-dependent enzyme highly expressed in key metabolic tissues, including the liver, adipose tissue, and the central nervous system. In these tissues, it reduces cortisone to the active hormone cortisone, which activates the glucocorticoid receptor. It belongs to the family of short-chain dehydrogenases. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.8 ★★★★★
Based on 17 reviews
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Product Reviews
★★★★★ 5
Nice AVR
Style: AVR-X1800H
Refurbished and looking like brand new. One small scratch on the side. It went in a cabinet and it's hidden. All paperwork seemed to be there. If I didn't know it looks like factory sealed . I've got an older Denon. Just wanted to move up to 7.2. Audyssey recognize a second sub. But neither did my older on. No problem with that though. Just FYI. After the original setup and minor tweaking it sounds fantastic for me. I haven't had it but a few days and I hope I get many years of enjoyment from this
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Reviewed in the United States on May 29, 2025
★★★★★ 5
Love it.
Style: AVR-X1800H
Love my Denon AV-1800 ....
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 9, 2025
★★★★★ 5
Best value
Style: AVR-X1800H
Great receiver ready to start awesome soung
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Reviewed in the United States on May 11, 2025
★★★★★ 5
Very good audio receiver for my tower speakers and record player set up
Style: Stereo Receiver, Configuration: Receiver
I have had my 1980s Roger sound lab CG 82 Tower speakers connected to a 1999 Technics audio receiver. I use this set up for Radio and my record player.
After many good years, the low end on my audio receiver gave out. I have had it serviced before, but I have owned it for five years and it is a 27-year-old piece of gear that has seen better days. ChatGPT led me to this receiver.
It took me less than five minutes to connect my speakers. What I love about this receiver is that it’s so simple to use. The radio signal is very good right out the box without an additional antenna. I was resistant to buying a Bluetooth receiver, but it connected to my iPad and my iPhone in seconds. Now I can play backing tracks through my Roger sound lab speaker speakers while I jam out on guitar.
No issues with sound quality or anything like that. Runs cool and quiet. If you are a total audio file, you could buy something more expensive. I am a musician and I listen to vinyl. Mostly jazz and classical, and this is fine for my needs. For most people running a record player, looking to modernize with a Bluetooth enabled receiver, this will suit your needs.
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Reviewed in the United States on April 15, 2026
★★★★★ 5
LOVE IT - Great Performance and Great Price
Style: Stereo Receiver, Configuration: Receiver, Style: Stereo Receiver, Configuration: Receiver
Speaking as a self-described "audiophile" (but a true audiophile with deep pockets would likely laugh), this was one of the best gifts I've given to myself in recent years (as I have a fairly large music library of nearly 600 albums of 1/3 vinyl, 2/3 CDs and some cassettes).
Bought mine back in March--used to be $200, but when I saw it for $150 any hesitation went out the window and I grabbed it. I have been in love with it ever since. Fabulous sound and power (do you really need more than 100 watts per channel?), great remote control (not used to that, believe it or not), and it lives up to Sony quality. Powers my vintage components perfectly:
> MCS Series 6202 Belt-Driven Turntable (had it since early 80s)
> Nacamichi BX-300 Cassette Deck
> Sony DVP-NS725P Disc Player (DVD but used mainly for CDs - audio-only connection)
> LG TV connected via optical-to-analog adapter
> KEF Reference Series 103/3 Stereo Speakers
Interesting observation about the way the ports are labeled on the back: what would traditionally be identified as a tape deck port (with IN/OUT) is just "Input 4," and while it's connected to my tape deck for playback, the REC-OUT is connected to a USB audio adapter to digitally record anything (mostly vinyl of course) to my laptop for mastering and iPod/digital library access.
The Bluetooth connectivity is awesome as well with powerful range - great for playing music from mobile devices (phone and iPod Classic with BT adapter). The front line-in 3.5 port, identified as a "Portable" is also a great feature.
At first, I was surprised that there was no input for optical, however since digital-to analog adapters are inexpensive and readily available, I think it's best to leave it that way. Would rather have the option, rather than an optical-only port that would otherwise go unused unless one had a component that required it.
HOWEVER, if I were to make any improvements, it would be to add a USB Out port (preferably on the front), for digital recording on a PC. However again, since adapters are inexpensive and accessible, and for the price I paid for this device, I ain't gonna complain. Front level analog balance, bass and treble controls would also be nice, but you have access to all that through the menu.
If you feel due for a simple, powerful and awesome stereo receiver/amplifier upgrade without breaking your monthly budget, this is a winner.
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Reviewed in the United States on December 9, 2023