Journal: PLoS ONE
Article Title: Context-dependent monoclonal antibodies against protein carbamidomethyl-cysteine
Figure Lengend Snippet: A series of purified proteins were treated in the presence or absence of the non-thiol reductant TCEP (2.5mM) with IAM or NEM (5mM). Proteins were then analyzed by Western blotting with 4E7, 52H11, or OX133. Selected CAM-cys labeled proteins reacted with 4E7 and 52H11. Equal loading of proteins was demonstrated by staining membranes with a non-selective protein stain (Revert, LI-COR). Proteins migrated at the expected molecular masses as follows (approximations): beta casein (27 kDa), BSA (65 kDa), BGN (40 kDa), DCN (38 kDa), COL1 (>250 kDa), COL4 (>250 kDa), JAG1 (170 kDa), IL17RC (75 kDa), LAMA2 (>180 kDa), vWF (260 kDa), TSP1 (150 kDa), TSP2 (155 kDa).
Article Snippet: Purified proteins were purchased from the following sources (all are human except where noted): Beta casein (bovine), Collagen I (gene:COL1), Collagen IV (Gene:COL4) (Sigma-Aldrich); Bovine serum albumin (gene:BSA) (bovine; New England BioLabs); Biglycan (gene:BGN), Decorin (gene:DCN), Jagged 1 (gene:JAG1), Interleukin 17 receptor C (gene:IL17RC), TSP1 and Thrombospondin-2 ((gene:TSP2) (R&D Systems)), Laminin subunit alpha 2 ((gene:LAMA2) (merosin; Millipore), vWF (Haematologic Technologies, Inc).
Techniques: Purification, Western Blot, Labeling, Staining