Journal: BMC Biology

Article Title: Proteomic analysis of blastema formation in regenerating axolotl limbs

doi: 10.1186/1741-7007-7-83

Figure Lengend Snippet: Highly regulated proteins

Article Snippet: Sections were then incubated over night with polyclonal anti-rabbit NOS1 (Biomol International LP, Plymouth Meeting, PA, USA) at 1:70 dilution, polyclonal anti-human fibronectin (Sigma, St Louis, MO, USA) at 1:400 dilution or monoclonal anti-α-actinin (Sigma) at 1:200 dilution, washed with blocking solution, incubated in the appropriate secondary antibody (goat anti-mouse AF488 or goat anti-rabbit AF568, Invitrogen, Carlsbad, CA, USA) for 40 min, washed with 1 × PBS and mounted with Vectashield mounting medium containing 4',6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Burlingame, CA, USA).

Techniques: Binding Assay, Translocation Assay

Journal: BMC Biology

Article Title: Proteomic analysis of blastema formation in regenerating axolotl limbs

doi: 10.1186/1741-7007-7-83

Figure Lengend Snippet: Immunostained sections of axolotl hindlimbs . Longitudinal sections of control (a, d, g) versus 1 day post amputation (dpa) (b, e, h) and 7 dpa (c, f, i) axolotl hindlimbs stained with primary antibodies to nitric oxide synthase 1 (NOS1) (a-c), fibronectin 1 (FN1) (d-f), α-actinin (ACTN) (g-i). Conjugated secondary antibodies were alexa-568 for fibronectin and NOS1, and alexa-488 for α-actinin. Nuclei were counterstained with 4',6-diamidino-2-phenylindole (DAPI). As expected from the proteomic data, fluorescence intensity of NOS1 showed a significant increase compared to control at 1 dpa, then decreased to a level slightly above control at 7 dpa. Fibronectin staining (red) at 1 and 7 dpa showed significant increases compared to controls, while α-actinin staining intensity (green) showed significant decreases.

Article Snippet: Sections were then incubated over night with polyclonal anti-rabbit NOS1 (Biomol International LP, Plymouth Meeting, PA, USA) at 1:70 dilution, polyclonal anti-human fibronectin (Sigma, St Louis, MO, USA) at 1:400 dilution or monoclonal anti-α-actinin (Sigma) at 1:200 dilution, washed with blocking solution, incubated in the appropriate secondary antibody (goat anti-mouse AF488 or goat anti-rabbit AF568, Invitrogen, Carlsbad, CA, USA) for 40 min, washed with 1 × PBS and mounted with Vectashield mounting medium containing 4',6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Burlingame, CA, USA).

Techniques: Control, Staining, Fluorescence

Journal: BMC Biology

Article Title: Proteomic analysis of blastema formation in regenerating axolotl limbs

doi: 10.1186/1741-7007-7-83

Figure Lengend Snippet: Liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS) versus densitometry measurements

Article Snippet: Sections were then incubated over night with polyclonal anti-rabbit NOS1 (Biomol International LP, Plymouth Meeting, PA, USA) at 1:70 dilution, polyclonal anti-human fibronectin (Sigma, St Louis, MO, USA) at 1:400 dilution or monoclonal anti-α-actinin (Sigma) at 1:200 dilution, washed with blocking solution, incubated in the appropriate secondary antibody (goat anti-mouse AF488 or goat anti-rabbit AF568, Invitrogen, Carlsbad, CA, USA) for 40 min, washed with 1 × PBS and mounted with Vectashield mounting medium containing 4',6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Burlingame, CA, USA).

Techniques: Chromatography, Comparison