Sulfo-NHS-SS-Biotin sodium

Sulfo-NHS-SS-biotin is a long-chain cleavable and cell-impermeant amine-reactive biotinylation reagent. Sulfo-NHS-SS-biotin can be used for the labeling and purifying of cell-surface protein[1].

Research Areas >> Others

Signaling Pathways >> Others >> Others

IC50: cell-surface protein[1]

Sulfo-NHS-SS-biotin is a cell-surface-labeling reagent, it is a negatively charged reagent and does not permeate cell membranes[1]. Sulfo-NHS-SS-biotin reacts with primary amines (NH2), such as lysine side-chains, or the amino-termini of polypeptides[1]. Sulfo-NHS-SS-biotin has a sulfonate group and prevents it from permeating cell membranes. Thr group’s cleavable spacer arm enables initially biotinylated proteins to be released from streptavidin affinity columns[1]. Sulfo-NHS-SS-biotin (1 mg/ml; 15 min) is applied in cells in monolayer culture (washed by ice-cold PBS). The biotinylation reactions are terminated with 100 mM glycine in PBS. After washing with PBS, cell extracts are prepared in RIPA buffer with protease inhibitor cocktail (HY-K0010). Biotinylated membrane proteins are precipitated with streptavidin-sepharose and proteins are eluted with SDS sample buffer[2].

[1]. Chia CP,et al. Phagocytosis in Dictyostelium discoideum is inhibited by antibodies directed primarily against common carbohydrate epitopes of a major cell-surface plasma membrane glycoprotein.Exp Cell Res. 1989 Mar;181(1):11-26.

[2]. Jo M, et al. Cell signaling by urokinase-type plasminogen activator receptor induces stem cell-like properties in breast cancer cells. Cancer Res. 2010 Nov 1;70(21):8948-58.