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ReadiPrep™ Plasma Membrane Protein Extraction Kit
Product key features
ReadiPrep™ Plasma Membrane Protein Extraction Kit enables selective isolation of plasma membrane proteins from cultured cells.
- Selective Membrane Protein Isolation: Effectively isolates plasma membrane proteins with minimal contamination from cytosolic or nuclear fractions.
- Streamlined Protocol: Complete membrane protein extraction without the need for ultracentrifugation or special equipment.
- High Compatibility: Suitable for downstream proteomics applications such as Western blotting, and ELISA.
Product description
The ReadiPrep™ Plasma Membrane Protein Extraction Kit is designed for efficient and reproducible isolation of plasma membrane proteins from cultured mammalian cells. Leveraging a detergent-based phase separation system, the kit selectively enriches plasma membrane proteins while minimizing contamination from cytosolic and nuclear components.
The protocol involves sequential lysis, temperature-controlled incubations, and phase separations that yield a fraction containing plasma membrane proteins, thus offering lower viscosity option compared to conventional plasma membrane protein extraction methods. The included buffers are optimized for maintaining protein integrity and compatibility with downstream analytical techniques such as Western blotting, proteomics, and functional assays. This kit is ideal for researchers studying membrane-bound receptors, transporters, or surface markers and suitable for both routine and high-throughput workflows.
Example protocol
AT A GLANCE
- Add ReadiPrep™ Lysis Buffer working solution to the cell pellet and incubate for 30 minutes on ice.
- Centrifuge the cells at 13000 X g for 5 minutes, and collect the supernatant in a new tube.
- Incubate the supernatant at 37°C for 10 minutes to separate the detergent phase.
- Centrifuge the cells at 13000 X g for 10 minutes at RT, and remove the aqueous phase (top layer).
- Add ReadiPrep™ Wash Buffer to the detergent phase and incubate on ice followed by 10 minutes at 37°C.
- Repeat the steps 4 and 5 to remove the aqueous phase for 3 times.
- Add ReadiPrep™ Dilution Buffer working solution to detergent phase and mix well.
Important: Check the buffers to see if they contain salt precipitates before use. If so, warm at room temperature or 37°C and shake the buffer until the salts are re-dissolved.
PREPARATION OF WORKING SOLUTION
Note: We recommend making the ReadiPrep™ Lysis Buffer working solution fresh as per need.
Note: We recommend making the ReadiPrep™™ Dilution Buffer working solution fresh as per need.
SAMPLE EXPERIMENTAL PROTOCOL
- Harvest cells and pellet the cells by centrifugation at 1000 rpm for 5 minutes.
- Resuspend the cells in 500 μL of ReadiPrep™ Lysis Buffer working solution and lyse the cells on ice for 30 minutes.
Note: It is recommended to use 80 to 90% confluence on a 10-cm plate. For optimal extraction, cell number must be optimized with indicated buffer volume. - Centrifuge the cells at 13000 X g for 5 minutes.
- Collect the supernatant into a new tube.
- Incubate the supernatant at 37°C for 10 minutes to separate the detergent phase.
- Centrifuge the cells at 13000 X g for 10 minutes at room temperature.
Note: It is necessary to perform the centrifugation step at room temperature to better separate the aqueous layer from the detergent phase. - Remove the aqueous phase (top layer) after the separation. The bottom layer is the detergent phase.
- Add equal volume (Approx. 300 to 350 μL) of ReadiPrep™ Wash Buffer (Component B) to the detergent phase.
- Incubate the mix on ice until you observe the solution getting cloudy.
- Incubate the mix at 37°C for 10 minutes to separate the detergent phase.
- Centrifuge at 13000 X g for 10 minutes at room temperature.
- Repeat Steps 7 to 11 for 3 to 4 times to completely remove the aqueous phase.
- Add equal volume of ReadiPrep™ Dilution Buffer working solution to the detergent phase. This mix contains the membrane proteins. Store at <-20 °C for downstream applications.
References
Authors: Moon, Youngsun and Lee, Yewon and Lee, Ho Jin and Byrne, Bernadette and Chae, Pil Seok
Journal: Current opinion in structural biology (2025): 103074
Authors: Suganuma, Nobuyasu and Saito, Nao and Yasukawa, Mio and Yamanaka, Takashi and Yamashita, Toshinari and Miyagi, Yohei and Saito, Aya and Hoshino, Daisuke
Journal: Anticancer research (2024): 5187-5192
Authors: Zhu, Hanlong and Zhao, Tianming and Zhao, Si and Yang, Suzhen and Jiang, Kang and Li, Shupei and Kang, Ying and Yang, Zhuoxin and Shen, Jiajia and Shen, Si and Tao, Hui and Xuan, Ji and Yang, Miaofang and Xu, Bing and Wang, Fangyu and Jiang, Mingzuo
Journal: Metabolism: clinical and experimental (2024): 155914
Authors: Smith, Baylee and Gaur, Deepika and Walker, Nathan and Walter, Isabella and Wohlever, Matthew L
Journal: bioRxiv : the preprint server for biology (2024)
Authors: Fresenius, Heidi L and Gaur, Deepika and Smith, Baylee and Acquaviva, Brian and Wohlever, Matthew L
Journal: bioRxiv : the preprint server for biology (2024)
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