Cell Surface Protein Isolation Kit (ab206998)
Key features and details
- Assay time: 3 hr
- Sample type: Adherent cells, Suspension cells
Overview
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Product name
Cell Surface Protein Isolation Kit -
Sample type
Adherent cells, Suspension cells -
Assay time
3h 00m -
Product overview
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 -
Notes
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.
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 10 tests DTT II 1 x 100µl Lysis Buffer VIII 1 x 6.5ml PBS Tablet 1 tablet Quenching Solution I 1 x 10ml Streptavidin Beads 1 x 1.5ml Sulfo-NHS-SS-Biotin 5 vials TBS Tablet 1 tablet Wash Buffer VII 1 x 12ml
Associated products
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Related Products
Images
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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).
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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.
Datasheets and documents
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SDS download
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Datasheet download
References (8)
ab206998 has been referenced in 8 publications.
- Chen Y et al. Amino acid starvation-induced LDLR trafficking accelerates lipoprotein endocytosis and LDL clearance. EMBO Rep 23:e53373 (2022). PubMed: 34994492
- Otsuji S et al. Clinical diversity and molecular mechanism of VPS35L-associated Ritscher-Schinzel syndrome. J Med Genet N/A:N/A (2022). PubMed: 36113987
- Chen YT et al. Endoplasmic reticulum protein TXNDC5 promotes renal fibrosis by enforcing TGF-ß signaling in kidney fibroblasts. J Clin Invest 131:N/A (2021). PubMed: 33465051
- He X et al. LYSMD3: A mammalian pattern recognition receptor for chitin. Cell Rep 36:109392 (2021). PubMed: 34289364
- Balic Z et al. Alternative splicing of the metalloprotease ADAMTS17 spacer regulates secretion and modulates autoproteolytic activity. FASEB J 35:e21310 (2021). PubMed: 33484187
- Chae CW et al. High glucose-mediated PICALM and mTORC1 modulate processing of amyloid precursor protein via endosomal abnormalities. Br J Pharmacol N/A:N/A (2020). PubMed: 32436237
- 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
- Wu X et al. Fucoidan elevates surface organic cation transporter 2 expression via upregulation of protein kinase A in uric acid nephropathy. Exp Ther Med 14:4153-4159 (2017). PubMed: 29104632