ubpbio/Linear Polyubiquitin Chain Capture Kit/J4450/1 Kit

来自 : 发布时间：2024-05-16

This kit is designed for enrichment of cellular proteins conjugated with linear polyubiquitin chains using GST-NEMO (UBAN). Polyubiquitinated proteins bound on GST-NEMO (UBAN) can be precipitated using glutathione resin and eluted by a buffer containing 10 mM glutathione. Polyubiquitinated proteins can be assayed by immunoblotting or mass spectrometry.

Additional Information

Product Name: Also Known As: Catalog No.: Size: Molecular Weight: Species: Source: Stock: Concentration: Quality Assurance: Storage: PDF Data Sheet: NCBI RefSeq: Image(s):

Linear Polyubiquitin Chain Capture Kit

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J4450

1 Kit

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Enzyme stocked in 20 mM Tris, 150 mM NaCl, 2 mM βME, 10% Glycerol

See tube label

Enzyme >95% by SDS-PAGE

Store at -80°C; avoid multiple freeze-thaw cycles

PDF datasheet, MSDS

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(Click image to enlarge)

Schematic of polyUb chains enrichment

Lane 1: GSTLane 2: GST-S5a (UIM)Lane 3: GST-TAB2 (NZF)Lane 4: GST-S5a (UIM) + GST-TAB2 (NZF)Lane 5: GST-NEMO (UBAN)According to procedures described above, 5 mg whole cell lysates from HeLa cells were mixed with 0.2 mg GST (lane 1), GST-S5a (UIM) (lane 2), GST-TAB2 (NZF) (lane 3), GST-S5a (UIM) + GST-TAB2 (NZF) (lane 4), or GST-NEMO (UBAN) (lane 5). 10% eluates were separated by SDS-PAGE, followed by immunoblotting of Ub, K48-specific Ub chains and GST. * No immunoreactivity was detected in GST-NEMO (UBAN) pulldown assays, presumably because there is little linear polyubiquitin chains in cells.

Shipping Method: Dry ice shipping References: N/A

Details

Description This kit is designed for enrichment of cellular proteins conjugated with linear polyubiquitin chains using GST-NEMO (UBAN). Polyubiquitinated proteins bound on GST-NEMO (UBAN) can be precipitated using glutathione resin and eluted by a buffer containing 10 mM glutathione. Polyubiquitinated proteins can be assayed by immunoblotting or mass spectrometry. Components • GST-NEMO (UBAN) (4 mg/ml)..................................0.5 ml • GST (4 mg/ml)...........................................................0.5 ml • Iodoacetamine (IAA)..................................................25 mg • Glutathione (reduced)...............................................100 mg• Glutathione Resin (75% slurry).................................1.0 ml Images:(Click image to enlarge) Schematic of polyUb chains enrichment Lane 1: GSTLane 2: GST-S5a (UIM)Lane 3: GST-TAB2 (NZF)Lane 4: GST-S5a (UIM) + GST-TAB2 (NZF)Lane 5: GST-NEMO (UBAN)According to procedures described above, 5 mg whole cell lysates from HeLa cells were mixed with 0.2 mg GST (lane 1), GST-S5a (UIM) (lane 2), GST-TAB2 (NZF) (lane 3), GST-S5a (UIM) + GST-TAB2 (NZF) (lane 4), or GST-NEMO (UBAN) (lane 5). 10% eluates were separated by SDS-PAGE, followed by immunoblotting of Ub, K48-specific Ub chains and GST. * No immunoreactivity was detected in GST-NEMO (UBAN) pulldown assays, presumably because there is little linear polyubiquitin chains in cells.