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Description
Human P-gp 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 permeability glycoprotein (P-gp) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of peroxidase (HRP) and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of permeability glycoprotein (P-gp) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Permeability Glycoprotein ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Permeability glycoprotein (P-gp or Pgp), also known as multidrug resistance protein 1 (MDR1), ATP-binding cassette subfamily B member 1 (ABCB1), or cluster of differentiation 243 (CD243), is an important protein of the cell membrane, encoded by the ABCB1 gene, which extracts many foreign substances from the cell. It is also a 170 kDa transmembrane glycoprotein, including 10-15 kDa of N-terminal glycosylation. The N-terminal half of the molecule contains six transmembrane domains, followed by a large cytoplasmic domain with an ATP binding site, and then a second part with six transmembrane domains and an ATP binding site, which has more than 65% amino acid similarity with the first half of the polypeptide. | |||||||||||||||||||||||||||||||||
| 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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4.9 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 5
Works perfectly with Macbook Air M2 - power, audio, video, sleep/wake, in clamshell mode!
My monitor has a USB-C video input that supports power delivery - so I was supposed to be able to charge my laptop with the same connection that goes to the monitor. Have had trouble finding an unpowered hub that actually supports all this without any hiccups, but according to all the specs I knew this was supposed to be possible.
But this one does! Now I can connect my apple laptop to my monitor, keyboard and mouse, and run in clamshell mode or with the laptop open, and it charges, wakes from sleep, does all the things I want. It also sends audio to the monitor (as it should). And I only need one USB-C connection to my laptop to do all this.
Note: I just got this hub so I don't know about longevity, and I haven't tested transfer speeds from all the ports on the hub, because that's not a big concern for me.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 25, 2026
★★★★★ 5
So far, so good. It's a USB-C hub, Anker has been doing USB hubs and power supplies for many years
I've always been impressed with Anker products, and great customer service! I do feel like they are starting to abandon the Android side of things recently, and the selection of cables has declined. USB-A cables seem to be discontinued, as well as USB-B.
I did have a really great CSR experience a while back. I had bought a Lithium Ion car battery jumpstarters, and had not used it for several years. I needed it one night, and the cables to connect to the battery had failed. Couldn't start my car. Even though the warranty was expired, and the product was even discontinued, they shipped me (at their cost) a replacement set of cables. Now a loyal customer.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 2, 2025
★★★★★ 1
Poor quality junk
Poor quality. Does not work as advertised. Do not buy this junk.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 14, 2026
★★★★★ 3
good on 16-inch MacBookPro and ThinkPad but not 13-inch MacBookPro; intermittent USB 3.2 connections
UPDATE 2025-NOV - lowering to 3-stars:
Found that only the 2nd USB-C from the right is truly USB 3.2 compliant. Using the same SSD and same USB-C 3.2 cable, only the 2nd port from the right connects at 10Gbps speeds for RTL9210B-chipset enclosure SSDs (and I have tested with 4 different ones). The other 2 ports on this Anker adapter are intermittent, sometimes they connect the SSD as USB 3.0 and sometimes it connects as USB 2.0, for the same SSDs. The odd thing (as possible workaround) is that if we use a USB-C to USB-A female adapter and then use a USB-A to USB-C cable to the same SSD, it now connects at USB 3.0 speed on any of the ports, but transfers only at 5Gbps throughput.
Another annoying thing is, this Anker adapter, even after the firmware upgrade, still intermittently "sleeps" or "goes offline" and disconnects from the laptop (does this on both my MacBook and ThinkPad). It does automatically reconnect after a few seconds, but very annoying, and can possibly corrupt drives that are connected when it happens.
ORIGINAL REVIEW - 4 stars:
USB-C hub is fast, able to transfer 5+ Gbps to SSD.
Video display quality is OK for 1440p (when it works).
DisplayPort video worked on ThinkPad and 16-inch MacBookPro, but not on 13-inch MacBookPro; connecting to a USB-C connected monitor (video worked without this Anker adapter).
USB-C cable length to host computer is long enough.
Worth the $30 price I paid ... yes; it served the purpose of having a USB-C hub while still being able to use the USB-C port for DisplayPort video; overall good design.
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Reviewed in the United States on September 10, 2025
★★★★★ 4
Video output does not contain power — caveats and findings for Switch users to whom this may concern
The product works with some caveats, which I initially tried to resolve with an ultimately unsuccessful firmware upgrade. (Anker was responsive via email, much appreciated.) It turns out the product was working to the best of its abilities this whole time, which I will now expound upon.
I expect that perhaps another 4-5 people in the world intended to use this in the ways that I did, namely with a Nintendo Switch as the source to various output devices. If you are one of these people who are interested in reading my experience connecting this hub with a Nintendo Switch, my findings are below (after a brief primer).
A Brief Primer
1. As you know, the Nintendo Switch can be coaxed to output Dp-Alt video if you can satisify a 39W+ handshake (15V, 2.6A or higher)
2. There are other adapters that will do this and output the video (with ample power) over USB-C, which are useful for powering AR glasses, portable monitors, or desktop USB-C monitors.
3. The limitation of a USB-C Dp-Alt adapter is that it does not also provide USB data ports like the Switch Dock itself, while many USB-C to HDMI dongles include USB-A ports. (But we don't want HDMI out, of course, because using USB-C we can power AR glasses or a portable monitor and send the video signal with one cable.)
4. I surmised that, by chaining a USB-C Dp Alt Adapter to this one (Anker A8340), I could have video output and three USB-C ports. Why do I want USB-C data ports? Since you are like me, you already know it is so I can connect a UAC1 DAC to output USB Audio and power thirsty headphones via a 4.4mm Pentacon balanced headphone jack. I also don't need to convince you of the merits of this versus using the Switch's built in 3.5mm headphone jack. Or say, connecting a wired controller because I don't care for input latency if I can avoid it.
My Findings
1. This hub can indeed be chained to a USB-C Dp-Alt adapter and pass through a video signal while also leaving three USB 3.2 data ports for wired devices (yes!)
2. However, as other reviews kindly mention, the video signal outputted by the A8340 does not contain power of any sort. So, if you are intending to plug this into a desktop USB-C monitor which already has/doesn't need power, then everything is awesome.
3. If you are trying to power AR glasses, this won't work unless you add a second USB-C DP-Alt adapter after it, and provide power that way. If you intend to convert the video signal to HDMI or DisplayPort via a USB-C to HDMI or DP cable, these won't work because there is no power for the inline conversion.
4. If you are using a USB-C travel monitor, this will work provided you directly power the monitor via its second USB-C power (if it has one).
So in summation, it's possible to get what you want, though you may find the victory pyrrhic. The perfect adapter (which I had hope this was, and may yet exist at a higher price point) basically takes a USB-C Dp-Alt adapter, adds USB-C data ports, and outputs USB-C + power to power AR glasses or a travel monitor.
This device nevertheless has its uses, such as the ability to connect multiple USB-C devices to the Switch in handheld mode and charge it, too.
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Reviewed in the United States on January 30, 2025
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