Key features and details
- Assay time: 3 hr
- Sample type: Adherent cells, Suspension cells
Product nameCell Surface Protein Isolation Kit
Sample typeAdherent cells, Suspension cells
Assay time3h 00m
Cell surface protein isolation kit (ab206998) enables the isolation of cell surface proteins for various downstream applications such as western blotting and other structural and functional studies. The cell surface protein isolation kit provides a simple and efficient method for the isolation of cell surface proteins.
In this method, cells are first labeled with Sulfo-NHS-SS-Biotin, an amine-reactive, thiol-cleavable, biotinylation reagent. Cells are subsequently lysed and the labeled cell surface proteins are isolated using streptavidin beads. The bound proteins are then released from beads by incubating with DTT solution. The biotinylation reagent is cell membrane-impermeable, with an extended spacer arm to reduce steric hindrances associated with streptavidin binding.
Cell surface protein isolation protocol summary:
- remove culture media and wash cells
- add biotin solution and incubate for 30 min
- add quenching solution and incubate for 5 min
- collect cells and spin at 500 g for 3 min
- wash cell pellet twice
- resuspend pellet in lysis buffer and incubate for 30 min
- spin at 10,000 g for 2 min and retain supernatant
- add supernatant to streptavidin beads and incubate for 1 hr
- spin at 800 g for 1 min
- wash beads twice and spin at 800 g
- add elution buffer and incubate for 30 min
- spin down beads and collect supernatant for analysis
This product is manufactured by BioVision, an Abcam company and was previously called K295 Cell Surface Protein Isolation Kit. K295-10 is the same size as the 10 test size of ab206998.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Storage instructionsStore at +4°C. Please refer to protocols.
Components Identifier 10 tests DTT Green 1 x 100µl Lysis Buffer Blue 1 x 6.5ml PBS Tablet WM 1 x 3 tablets Quenching Solution NM 1 x 10ml Streptavidin Beads Violet 1 x 1.5ml Sulfo-NHS-SS-Biotin Amber 5 vials TBS Tablet NM 1 tablet Wash Buffer WM 1 x 12ml
western blot analysis of cell surface proteins isolated using ab206998 Cell surface proteins were isolated from HeLa cells in the presence (+) and absence (-) of Sulfo-NHS-SS-Biotin. The blots were probed with antibodies to Endoplasmic Growth Factor Receptor (EGFR), a plasma membrane marker (top) and Calnexin, a cytosolic marker (bottom).
western blot analysis of isolated cell surface proteins HeLa cells were used to isolate the cell surface proteins, in the presence (+) and absence (-) of Sulfo-NHS-SS-Biotin. (b) Integrin β1 and Hsp90 as plasma membrane and cytosolic markers, respectively. Isolation was performed following the kit protocol.
ab206998 has been referenced in 2 publications.
- Balic Z et al. Alternative splicing of the metalloprotease ADAMTS17 spacer regulates secretion and modulates autoproteolytic activity. FASEB J 35:e21310 (2021). PubMed: 33484187
- Mishra S et al. The protein secretion modulator TMED9 drives CNIH4/TGFa/GLI signaling opposing TMED3-WNT-TCF to promote colon cancer metastases. Oncogene 38:5817-5837 (2019). PubMed: 31253868