Tyrosine Kinase Receptor A (TrkA), Mouse Monoclonal Antibody

Western blot of TrkA in rat PC12 cell lysates (64 µg/lane): SDS-PAGE: denatured, non-reduced; Transfer: Tris-Glycine buffer; Membrane: nitrocellulose (0.45 um); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: overnight at 4°C (3 µg/mL); Secondary antibody: anti-mouse-HRP (1/6000) 2 hours at RT; Detection: Chemiluminiscence. M-1719-100 detects membrane-associated TrkA full-length protein at approximately 140 kDa (gp140TrkA). Predicted MW of rat TrkA based on amino acid sequence: 85 kDa. The observed MW differs due to post-translational modification, mainly glycosylation.

Western blot of TrkA in rat PC12 cell lysates (80 µg/lane). M-1719-100 detects the intracellular glycosylated immature form of TrkA at 110 kDa (gp140TrkA), and an intracellular cleavage product at 41 kDa (p41, see Cabrera et al., 1995; Rossi et al., 2002). These bands are TrkA-specific, because detection by M-1719-100 is abolished after pre-incubation with blocking peptide. SDS-PAGE: denatured, non-reduced; Transfer: Tris-Glycine buffer; Membrane: nitrocellulose (0.45 µm); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: overnight at 4°C (3 µg/mL); Secondary antibody: anti-mouse-HRP (1/6000) 2 hours at RT; Detection: Chemiluminiscence.

Western blot of TrkA in rat PC12 cell lysates (30 µg/lane). M-1719-100 detects membrane-associated TrkA full-length protein at 140 kDa (gp140TrkA), the glycosylated immature form of TrkA at 110 kDa (gp110TrkA) and an intracellular cleavage product at 41 kDa (p41, see Cabrera et al., 1995; Rossi et al., 2002). SDS-PAGE: denatured, reduced and non-reduced; Transfer: Tris-Glycine buffer; Membrane: nitrocellulose (0.45 µm); Blocking: 5% skim milk in TBST, 2 hours at RT; Primary antibody: overnight at 4°C (3 µg/mL); Secondary antibody: anti-mouse-HRP (1/6000) 2 hours at RT; Detection: Chemiluminiscence.

Immunofluorescence analysis of TrkA expression in human SH-SY5Y cells. Fixed (4% formaldehyde), permeabilized, and blocked (10% normal horse serum, 0.1% Triton X100) SH-SY5Y cells were incubated with TrkA antibody M-1719-100 (2 µg/mL, green) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-mouse-CF488A antibody (4 µg/mL, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue). Magnification: 100x.

Immunofluorescence analysis of TrkA expression in human DU145 prostate cancer cells. Fixed (4% formaldehyde), permeabilized, and blocked (10% normal horse serum, 0.1% Triton X100) DU145 cells were incubated with TrkA antibody M-1719-100 (2 µg/mL, green) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-mouse-CF488A antibody (4 µg/mL, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue). Magnification: 100x.

TrkA staining in fixed and permeabilized embryonic rat DRG neurons by Immunocytochemistry. TrkA-IR is observed in axonal projections (white arrows) and cell body (membrane and/or cytoplasmic compartments, yellow arrow), consistent with expected cellular localization. Primary antibody concentration: 2 µg/mL; Magnification: 32X. Image courtesy of QBM Cell Science.