polyvinylidene fluoride membranes pvdf  (Bio-Rad)

Journal: The Plant Cell

Article Title: The Reverse Transcriptase/RNA Maturase Protein MatR Is Required for the Splicing of Various Group II Introns in Brassicaceae Mitochondria

doi: 10.1105/tpc.16.00398

Figure Lengend Snippet: Relative Accumulation of MatR and Various Mitochondrial Proteins during Arabidopsis Seed Germination. (A) ). Detection was performed by chemiluminescence with the Image Quant LAS4000 mini analyzer (GE Healthcare). The intensities of protein signals in (A) and (B) ). (B) -PAGE analysis of the respiratory chain complexes during seed germination in Arabidopsis. Crude membrane fractions obtained from dry seeds, imbibed seeds, and mature Arabidopsis seedlings were solubilized with DDM ( n ). For immunodetections, the proteins were transferred from the native gels onto a PVDF membrane (Bio-Rad) in cathode buffer for 15 h at 40 mA, using the Bio-Rad mini transblot cell. The membranes were distained with ethanol before probing with specific antibodies, as indicated below each blot. Arrows indicate the native respiratory complexes, CI (∼1000 kD), CIII (dimer, ∼500 kD), CIV (∼220 kD), and CV (∼600 kD), in Arabidopsis mitochondria. Please note, in (B) , the original COX2 blot has been modified, i.e., the lane corresponding of the mature leaves (M) was cut from the right side of the blot and pasted to the other side (marked with dotted line). No other changes have been made to the original figure.

Article Snippet: For nondenaturing-PAGE-protein gel blotting, the gel was transferred to a PVDF membrane (Bio-Rad) in Cathode buffer (50 mM Tricine and 15 mM Bis-Tris-HCl, pH 7.0) for 16 h at 4°C (constant current of 40 mA).

Techniques: Polyacrylamide Gel Electrophoresis, Modification

Journal: Iranian Biomedical Journal

Article Title: Expression Analysis of RNA-Binding Motif Gene on Y Chromosome (RBMY) Protein Isoforms in Testis Tissue and a Testicular Germ Cell Cancer-Derived Cell Line (NT2)

doi: 10.6091/ibj.1148.2013

Figure Lengend Snippet: RBMY antibody confirmation by using Western-blot analysis of recombinant RBMY. Two concentrations (100 and 10 ng) of each recombinant protein were loaded on SDS-PAGE and after separation were transferred to PVDF membrane. Blots were exposed to X-ray films

Article Snippet: Total protein extract (40 µg) from each sample was separated on 12% SDS-polyacrylamide gels for 120 min at 100 V and transferred to PVDF membrane (Bio-Rad, USA) by a wet transfer system (Bio-Rad, USA) at 20 V overnight.

Techniques: Western Blot, Recombinant, SDS Page

Journal: Bioorganic & medicinal chemistry

Article Title: Bivalent ligands incorporating curcumin and diosgenin as multifunctional compounds against Alzheimer’s disease

doi: 10.1016/j.bmc.2015.10.032

Figure Lengend Snippet: Compounds 33 and 38 bind Aβ and reduce AβO formation, but have no effect on Aβ production. (A) Representative western blot. Cells were treated were lysed, and proteins were separated by SDS-PAGE. After transfer to a PVDF membrane, blots were probed with the 6E10 antibody. (B) Quantification of total Aβ oligomers from western blotting. Error bars represent SEM. (n=6; ** p

Article Snippet: Equal amounts of protein (20.0 μg) were separated by SDS-PAGE on a Tris-Tricine gel (Bio-Rad) and transferred onto a PVDF membrane (Bio-Rad).

Techniques: Western Blot, SDS Page

Journal: PLoS ONE

Article Title: Control of organelle gene expression by the mitochondrial transcription termination factor mTERF22 in Arabidopsis thaliana plants

doi: 10.1371/journal.pone.0201631

Figure Lengend Snippet: Relative accumulation of organellar proteins in wild-type and mterf22 plants. (A) Immunoblot analyses of wild-type plants and mterf22-1 mutant line. For the quantification of the relative abundances of organellar proteins in mterf22 plants, different amounts of total mitochondrial proteins extracted from wild-type plants were loaded and separated by SDS-PAGE. The blots were probed with polyclonal antibodies raised to different organellar proteins, as indicated in each panel. Detection was carried out by chemiluminescence assays after incubation with HRP-conjugated secondary antibody. (B) BN-PAGE of crude mitochondria preparations was performed according to the method described in [ 57 ]. Crude mitochondria preparations, obtained from 3-week-old Arabidopsis seedlings, were solubilized with DDM [1.5% (w/v)] and the organellar complexes were resolved by BN-PAGE. For immunodetection, proteins were transferred from the native gels onto a PVDF membrane and were probed with specific antibodies ( S2 Table ), as indicated below each blot. Arrows indicate to the native complexes I (~1,000 kDa), III (dimer, ~500 kDa), IV (~220 kDa) and V (~600 kDa). The asterisk in the CA2 panel indicates to the presence of a 700 ~ 800 kDa band, which may corresponds to a complex I assembly intermediate. Hybridization signals were analyzed by chemiluminescence assays after incubation with HRP-conjugated secondary antibody. The intensities of protein signals in panels ‘A’ and ‘B’ using ImageJ software [ 90 ].

Article Snippet: For immunoblotting of non-denaturing PAGE, the proteins were transferred from the gel onto a PVDF membrane (Bio-Rad) in Cathode buffer (50 mM Tricine, 15 mM Bis-Tris-HCl, pH 7.0) for 16 h at 4°C at constant current of 40 mA.

Techniques: Mutagenesis, SDS Page, Incubation, Polyacrylamide Gel Electrophoresis, Immunodetection, Hybridization, Software