Product Name Anti-6-Nitrotryptophan Mouse Monoclonal Antibody (56619)
Description Anti-6-Nitrotryptophan Mouse Monoclonal Antibody
Species Reactivity Species Independent
Clone ID 2D12
Immunogen Synthetic 6-Nitrotryptophan modified Keyhole Limpet Hemocyanin (KLH).
Concentration 1 mg/mL
Formulation PBS pH 7.4, 50% glycerol, 0.09% Sodium azide
Buffer Formulation Phosphate Buffered Saline
Buffer pH pH 7.4
Buffer Anti-Microbial 0.09% Sodium azide
Buffer Cryopreservative 50% glycerol
Purification Protein G Purified
Background 6-nitrotryptophan is a major nitrated product of tryptophan residues in proteins reacted with reactive nitrogen species (RNS) (1). Superoxide (O2-) and nitric oxide (NO) react to form peroxynitrous acid, which subsequently nitrates tryptophan residues, modulating protein functions (2). Tryptophan residues are more likely to be nitrated when the indole ring is exposed to the surface of proteins (1).
Specificity Specific for 6-Nitrotyrptophan modified proteins. Doss not cross-react with 7-Ketocholoesterol modified proteins.
Target Name Nitrotryptophan
Target ID Nitrotryptophan
Alternative Names 6-Nitrotryptophan, Nitrotryptophan (NW), Nitrotryptophan, NW, 6-NO2-Trp
PubChem ID 3246256
Research Areas Cell Signaling | Post-translational Modifications | Nitration
Description Western Blot analysis of 6-Nitrotryptophan-BSA Conjugate showing detection of 67 kDa Nitrotryptophan protein using Mouse Anti-Nitrotryptophan Monoclonal Antibody, Clone 2D12 . Lane 1: Molecular Weight Ladder (MW). Lane 2: BSA (0.5 µg). Lane 3: BSA (1 µg). Lane 4: 6-Nitrotryptophan-BSA (0.5 µg). Lane 5: 6-Nitrotryptophan-BSA (1 µg). Lane 6: 7-Ketocholesterol-BSA (0.5 µg). Lane 7: 7-Ketocholesterol-BSA (1 µg). Block: 5% Skim Milk in TBST. Primary Antibody: Mouse Anti-Nitrotryptophan Monoclonal Antibody at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Mouse IgG: HRP at 1:2000 for 60 min at RT. Color Development: Luminol for 1 min. Predicted/Observed Size: 67 kDa.
Description Flow Cytometry analysis using Mouse Anti-Nitrotryptophan Monoclonal Antibody, Clone 2D12 . Tissue: Neuroblastoma cells (SH-SY5Y). Species: Human. Fixation: 90% Methanol. Primary Antibody: Mouse Anti-Nitrotryptophan Monoclonal Antibody at 1:50 for 30 min on ice. Secondary Antibody: Goat Anti-Mouse: PE at 1:100 for 20 min at RT. Isotype Control: Non Specific IgG. Cells were subject to oxidative stress by treating with 250 µM H2O2 for 24 hours.
Description Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Nitrotryptophan Monoclonal Antibody, Clone 2D12 . Tissue: Embryonic kidney epithelial cell line (HEK293). Species: Human. Fixation: 5% Formaldehyde for 5 min. Primary Antibody: Mouse Anti-Nitrotryptophan Monoclonal Antibody at 1:50 for 30-60 min at RT. Secondary Antibody: Goat Anti-Mouse Alexa Fluor 488 at 1:1500 for 30-60 min at RT. Counterstain: Phalloidin Alexa Fluor 633 F-Actin stain; DAPI (blue) nuclear stain at 1:250, 1:50000 for 30-60 min at RT. Localization: Cytoplasmic. Magnification: 20X (2X Zoom). (A) DAPI (blue) nuclear stain. (B) Phalloidin Alexa Fluor 633 F-Actin stain. (C) Nitrotryptophan Antibody. (D) Composite. Courtesy of: Dr. Robert Burke, University of Victoria.
Storage This antibody is stable for at least one (1) year at -20°C. Avoid multiple freeze-thaw cycles.
Dilution Instructions Dilute in PBS or medium which is identical to that used in the assay system.
Application Instructions WB (1:1000); ICC/IF (1:50); FACS (1:50); FCM (1:50); ELISA (1:1000); optimal dilutions for assays should be determined by the user.