primary antibodies against gb3  (AMS Biotechnology)

Journal: Journal of nanobiotechnology

Article Title: Ceria-Zirconia nanoparticles reduce intracellular globotriaosylceramide accumulation and attenuate kidney injury by enhancing the autophagy flux in cellular and animal models of Fabry disease.

doi: 10.1186/s12951-022-01318-8

Figure Lengend Snippet: Fig. 3 Gb3 detection in α-GLA knockdown HK-2 cells and human podocytes at baseline and under PEG-CZNPs treatment. A–D HK-2 cell analysis A Representative confocal immunofluorescence microscopy images of Gb3 (green), with DAPI counterstaining (blue), in control (siRNA) and α-GLA deficient HK-2 cells (siGLA), with or without exposure to PEG-CZNPs. B Quantification of the fluorescence intensities in the confocal microscope images using ImageJ software. C Representative TEM images of control (siRNA) and α-GLA deficient HK-2 cells (siGLA), with or without exposure to PEG-CZNPs. D Quantitative analysis of Gb3 vesicles in control (siRNA) and α-GLA deficient HK-2 cells (siGLA), with or without exposure to PEG-CZNPs. E–I Human podocyte analysis E Representative confocal immunofluorescence microscopy images of Gb3 (green), with DAPI counterstaining (blue), in control (siRNA) and α-GLA deficient human podocytes (siGLA) with or without exposure to PEG-CZNPs. F Quantification of the fluorescence intensity of confocal microscope images using ImageJ software. G Representative TEM images of control (siRNA) and α-GLA deficient human podocytes (siGLA) with or without exposure to PEG-CZNPs. H Quantitative analysis of Gb3 vesicles from TEM images. I Quantification of the total Gb3 content using by LC–MS/MS data and the Student’s t test. Data shown are the mean ± SD; *P < 0.05, **P < 0.01, and ***P < 0.001 versus control; #P < 0.05, ##P < 0.01, and ###P < 0.001 versus siGLA knockdown alone

Article Snippet: IFA was also performed Using primary antibodies against Gb3 (Amsbio, Cambridge, MA), synaptopodin (Sigma Aldrich) and LC3B (Sigma Aldrich).

Techniques: Knockdown, Cell Analysis, Immunofluorescence, Microscopy, Control, Fluorescence, Software, Liquid Chromatography with Mass Spectroscopy

Journal: Journal of nanobiotechnology

Article Title: Ceria-Zirconia nanoparticles reduce intracellular globotriaosylceramide accumulation and attenuate kidney injury by enhancing the autophagy flux in cellular and animal models of Fabry disease.

doi: 10.1186/s12951-022-01318-8

Figure Lengend Snippet: Fig. 12 Schematic illustrations of the PEG-CZNP induced attenuation of kidney injury from FD. This occurs via the enhancement of the autophagy flux combined with the activation of TFEB, restoration of AKT/mTOR signaling, and antioxidant effects. PEG-CZNPs thereby alleviate inflammatory and fibrosis pathways and attenuate Gb3-mediated kidney injury

Article Snippet: IFA was also performed Using primary antibodies against Gb3 (Amsbio, Cambridge, MA), synaptopodin (Sigma Aldrich) and LC3B (Sigma Aldrich).

Techniques: Activation Assay

Journal:

Article Title: Defective Laminin 5 Processing in Cylindroma Cells

doi:

Figure Lengend Snippet: Indirect immunofluorescence staining of laminins and nidogen in cylindromas (A–C) and at the dermal-epidermal junction (D–F). Cryosections (5 μm) of tissues were incubated with mouse monoclonal antibodies BM165 (A and D) or 4C7 (B and E) against the human laminin α3 and α5 chains, respectively, or with a polyclonal serum against nidogen 1 (C and F). Mouse or rabbit immunoglobulins were detected with Cy3-conjugated secondary antibodies. For each different staining, the pictures corresponding to the cylindroma islands (A–C) or to the dermal-epidermal junction are from the same tissue section. Immunoreactivity corresponding to the laminin α3 chain is observed in the whole thickness of the material surrounding the tumor islets (A). By contrast, immunoreactivity corresponding to the laminin α5 chain (B) or to nidogen (C) is mainly restricted to the inner face of the basement membrane outlining the cylindromas, and for α5 to some intranodular deposits.

Article Snippet: For indirect immunofluorescence staining primary mouse monoclonal antibodies against laminin chains included BM165 against α3, GB3 against γ2 (Seralab), and 4C7 against native α5 chain (Life Technologies).

Techniques: Immunofluorescence, Staining, Incubation, Bioprocessing, Membrane

Journal:

Article Title: Defective Laminin 5 Processing in Cylindroma Cells

doi:

Figure Lengend Snippet: Altered processing of laminin 5 in primary cultures of cylindroma cells. The culture medium of cylindroma cells (Cy) and of primary human keratinocytes (Ke) was mixed 1:1 with Laemmli buffer and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions on 4 to 10% acrylamide gels (50 μl/lane). After transfer to nitrocellulose membranes, the blots were incubated with mouse monoclonal antibodies BM165 (A) or D4B5 (B) against the human laminin α3 or γ2 chains, respectively. The blot shown in B was extensively washed and reblotted with a goat antibody against a carboxy-terminal peptide of the laminin β3 chain (C). Bound antibodies were detected using horseradish peroxidase-conjugated secondary antibodies followed by enhanced chemiluminescence. Molecular weight markers are indicated at the right of the blots. Arrowheads at the left of the blots indicate the migration mobility of the different laminin chains. The relative optical density of the bands corresponding to differently processed forms of the laminin α3A and γ2 chains was measured by scanning densitometry. Each column represents the relative percentage of the processed (dots) and unprocessed (stripes) forms of the laminin α3A (D) and γ2 (E) chains shown in blots A and B, respectively.

Article Snippet: For indirect immunofluorescence staining primary mouse monoclonal antibodies against laminin chains included BM165 against α3, GB3 against γ2 (Seralab), and 4C7 against native α5 chain (Life Technologies).

Techniques: Polyacrylamide Gel Electrophoresis, Incubation, Bioprocessing, Molecular Weight, Migration