Journal: Journal of Biological Chemistry

Article Title: Subcellular Localization of the Aryl Hydrocarbon Receptor Is Modulated by the Immunophilin Homolog Hepatitis B Virus X-associated Protein 2

doi: 10.1074/jbc.m006873200

Figure Lengend Snippet: FIG. 5. Examination of AhR localization and AhR/XAP2 stoichiometry in Hepa-1 cells. A, laser scanning confocal micrographs of AhR in control and 1 nM TCDD-treated Hepa-1 cells visualized with anti-AhR MAb RPT9 and goat anti-mouse IgG conjugated to lissamine-rhodamine sulfonyl chloride, Me2SO (i), 30 min (ii), 1 h (iii), 2 h (iv). B, relative amount of AhR in Hepa-1 cytosolic fraction compared with total AhR (cytosolic 1 nuclear). Cells were treated with 1 nM TCDD for 2 h, harvested, homogenized, and centrifuged to obtain nuclear pellet and crude cytosol (respun to obtain cytoplasm). Nuclear pellet was extracted with MENG (1500 mM NaCl), and soluble extract collected. Aliquots of cytosolic and nuclear extracts were resolved by TSDS-PAGE, blotted to PVDF membrane, and visualized using the RPT1/biotin-goat anti-mouse IgG/125I-labeled streptavidin system. C, sedimentation profile of AhR from cytosolic extract of Hepa-1 cells treated with 1 nM TCDD for 2 h. Extracts were applied to 10–30% sucrose gradient in MENG and fractionated. Fractions were acetone precipitated and analyzed by SDS-PAGE, visualized as in panel B, and bands excised and quantitated in a gamma counter. D, ratios of AhR to XAP2 in Hepa-1 cells in cytosolic extracts (cytosol) and in the immunoprecipitated 9 S AhR (complex).

Article Snippet: The membrane was then analyzed by Western blot with RPT1 (anti-AhR MAb) and anti-ARA9 (anti-XAP2 MAb; Novus Biologicals) primary antibodies, and 125I-labeled goat anti-mouse IgG (PerkinElmer Life Sciences) secondary Ab.

Techniques: Control, Membrane, Labeling, Sedimentation, SDS Page, Immunoprecipitation

Journal: Separations

Article Title: Alternative Method for HDL and Exosome Isolation with Small Serum Volumes and Their Characterizations

doi: 10.3390/separations8110204

Figure Lengend Snippet: Figure 2. SDS-PAGE and Western blotting of HDLs isolated from different sample volumes. A protein ladder reference (Vivantis Technologies, Shah Alam, Selangor Darul Ehsan, Malaysia, cat no. PR0602) had 3 reference bands (10.5, 42, and 95 kDa) coupled with a blue dye for easy identification (A). Based on Coomassie blue G250 staining of SDS-PAGE, major bands were identified approximately at 29, 51, and 70 kDa in both HDL200 and HDL500 (B). A single band was identified at about 29 kDa corresponding to ApoA-1 (C). M = Marker of protein molecular weight.

Article Snippet: The enhanced luminol-based chemiluminescent (ECL) Western blotting reagent (cat no. RPN2232) was a product from GE Healthcare, Chalfont St Giles, Buckinghamshire, UK.

Techniques: SDS Page, Western Blot, Isolation, Staining, Marker, Molecular Weight

Journal: Separations

Article Title: Alternative Method for HDL and Exosome Isolation with Small Serum Volumes and Their Characterizations

doi: 10.3390/separations8110204

Figure Lengend Snippet: Figure 6. The protein pattern and biomarker of the exosomes isolated from different sample volumes. A protein ladder reference (Vivantis Technologies, Shah Alam, Selangor Darul Ehsan, Malaysia, cat no. PR0623) had 2 reference bands (25 and 72 kDa) coupled with blue chromophore for easy identification (A). SDS-PAGE stained with Coomassie blue G250 showed multiple bands with the same patterns (B). The Western blot results showed a single band of TSG101 at about 48.9 kDa exhibiting the exosome biomarker (C). Thirty micrograms of protein were loaded for both SDS-PAGE and the Western blot. All experiments above were run with a reducing agent. M = Marker of protein molecular weight.

Article Snippet: The enhanced luminol-based chemiluminescent (ECL) Western blotting reagent (cat no. RPN2232) was a product from GE Healthcare, Chalfont St Giles, Buckinghamshire, UK.

Techniques: Biomarker Discovery, Isolation, SDS Page, Staining, Western Blot, Marker, Molecular Weight

Journal: iScience

Article Title: Nucleation and dissolution mechanism underlying amyotrophic lateral sclerosis/frontotemporal lobar dementia-linked fused in sarcoma condensates

doi: 10.1016/j.isci.2023.106537

Figure Lengend Snippet:

Article Snippet: 10x Tris/Glycine Buffer for Western Blots and Native Gels , Bio-Rad , 1610772.

Techniques: Virus, Recombinant, Protease Inhibitor, Lysis, Western Blot, Software, Pore Size, Plasmid Preparation