mouse monoclonal antibody against green fluorescent protein  (Santa Cruz Biotechnology)

Journal: PLoS ONE

Article Title: Functional Interactions between BM88/Cend1, Ran-Binding Protein M and Dyrk1B Kinase Affect Cyclin D1 Levels and Cell Cycle Progression/Exit in Mouse Neuroblastoma Cells

doi: 10.1371/journal.pone.0082172

Figure Lengend Snippet: ( a ) Neuro 2a cells were transiently single- and double-transfected with: Dyrk1B, pCAG-Cend1-IRES-GFP, RanBPM, RanBPM and Dyrk1B, RanBPM and pCAG-Cend1-IRES-GFP expression plasmids, as indicated. Cells were allowed 16 h for protein expression and were then induced to differentiate by treatment with 20 μM RA/ 2% FCS for 48 h after which they were fixed and immunofluorescently labeled for Dyrk1B, Cend1, RanBPM and the neuronal differentiation marker βIII-tubulin (Tuj1 antibody). Note that Cend1 is co-expressed with GFP (v, vi) which serves for visualization of Cend1-positive cells (vi, xviii). Scale bar: 40 μm. ( b ) Quantification of the mean neurite length in single and double-transfected cells (as indicated) as well as in non-transfected cells in each group. Different groups were compared by one-way ANOVA followed by Student's t-test. ***Student’s t-test: p<0.001, **: p<0.01, *: p<0.05, n= 3. Error bars represent SEM. #: represents statistically significant differences between transfected and non-transfected cells in the same group, while *: represents statistically significant differences among different groups.

Article Snippet: Other antibodies also used in this study were rabbit polyclonal antibodies against β-tubulin (sc-9104), α-actin and β-actin (sc-1615), mouse monoclonal antibody against βIII-tubulin (Tuj1; Covance, MMS-435P), mouse monoclonal antibody against green fluorescent protein (GFP) (Invitrogen), rabbit polyclonal against phospho-histone 3 (PH3) (Upstate) and goat polyclonal antibody against Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Santa Cruz, sc-20357).

Techniques: Transfection, Expressing, Labeling, Marker

Journal: The New phytologist

Article Title: UBC18 mediates ERF1 degradation under light-dark cycles.

doi: 10.1111/nph.14272

Figure Lengend Snippet: Fig. 1 ERF1 protein levels under dark incubation. (a) The upper panel shows that green fluorescent protein (GFP) signals in 4- d-old roots of 35S:ERF1-GFP transgenic Arabidopsis thaliana plants become weaker after a 6 h dark incubation (WL 4h to D). The lower panel shows that GFP signals remain present under continuous light conditions (cWL). Bars, 10 lm. (b–d) Western blot analyses of ERF1 protein (by GFP antibody) in 15-d-old 35S:ERF1-GFP plants. The plants grown on MS were first illuminated for 4 h and then incubated for up to 8 h in the dark (WL 4h to D, b, d) or first incubated in the dark for 8 h and then illuminated for up to 4 h (D8 to WL, c). (d) Western blot analysis of ERF1 protein in the 35S:ERF1-GFP transgenic plants during dark incubation treated with MG132. Bars indicate SE. (e) ERF1 expression in the 35S:ERF1-GFP transgenic plants during dark incubation.

Article Snippet: Monoclonal mouse antibody against green fluorescent protein (GFP) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Incubation, Transgenic Assay, Western Blot, Expressing

Journal: The New phytologist

Article Title: UBC18 mediates ERF1 degradation under light-dark cycles.

doi: 10.1111/nph.14272

Figure Lengend Snippet: Fig. 7 UBC18 enzyme activity is required for degradation of ERF1. Immunoblot analyses of the expression of ERF1 when coexpressed with the functional UBC18WT or catalytically defective UBC18C99A in Nicotiana benthamiana leaves. Coinfiltration of the green fluorescent protein (GFP) was used as an internal control. Note that ‘+++’ and ‘++’ denote five- and 2.5-fold increases in the volume of infiltration relative to ‘+’, respectively. –, not added. Experiments were repeated three times with similar results, and a representative experiment is shown.

Article Snippet: Monoclonal mouse antibody against green fluorescent protein (GFP) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Activity Assay, Western Blot, Expressing, Functional Assay, Control

Journal: The New phytologist

Article Title: UBC18 mediates ERF1 degradation under light-dark cycles.

doi: 10.1111/nph.14272

Figure Lengend Snippet: Fig. 8 UBC18 mediated the ubiquitination of ERF1. Immunoblot analysis by anti-GFP or anti-ubiquitin antibody following immunoprecipitation (IP) of total proteins. Total proteins were isolated from wild-type (WT), ubc18- 1, and ubc18-2 Arabidopsis thaliana seedlings transfected with mock, green fluorescent protein (GFP) only, and ERF1-GFP using the AGROBEST system. Arrows indicate the ERF1-GFP fusion protein. MW, molecular weight.

Article Snippet: Monoclonal mouse antibody against green fluorescent protein (GFP) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Ubiquitin Proteomics, Western Blot, Immunoprecipitation, Isolation, Transfection, Molecular Weight