Antibody was affinity purified using an epitope specific to DJ-1 immobilized on solid support. The epitope recognized by A300-744A maps to a region between residue 140 and the C-terminus (residue 189) of human DJ-1 (PARK7, Parkinson Disease 7) using the numbering given in entry NP_009193.2 (GeneID 11315 ). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
DJ1, DJ-1, epididymis secretory sperm binding protein Li 67p, GATD2, HEL-S-67p, maillard deglycase, oncogene DJ1, Parkinson disease (autosomal recessive, early onset) 7, Parkinson disease protein 7, parkinson protein 7, Parkinsonism-associated deglycase,
between 140 and C-term
2 - 8°C
Detection of human DJ-1 by western blot. Samples: Whole cell lysate (50 µg) from HeLa and HEK293T cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-DJ-1 antibody A300-744A (lot A300-744A-1) used for WB at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.
Detection of human and mouse DJ-1 by immunohistochemistry. Sample: FFPE section of human breast carcinoma (left) and mouse teratoma (right). Antibody: Affinity purified rabbit anti-DJ-1 (Cat. No. A300-744A Lot1) used at a dilution of 1:1,000 (0.2µg/ml). Detection: DAB
Detection of human DJ-1 by western blot and immunoprecipitation/Mass Spectrometry. Samples: A) Whole cell lysate from HeLa (20 and 4 µg for WB), MCF-7 (7; 20µg) and HEK293T (T; 20µg) cells. B) Cytosolic and nuclear extract (10 mg/IP) from HeLa S3 cells. Antibodies: Affinity purified rabbit anti-DJ-1 antibody (Cat.No. A300-744A) used for WB at 0.1 µg/ml (A) and for IP at 0.7 µg/mg lysate (B). Detection: Chemiluminescence with an exposure time of 5 sec (A) and vMALDI-LTQ and LC-MS/MS of Coomassie stained bands (B).