rabbit polyclonal anti lamp1 antibody  (Santa Cruz Biotechnology)

Journal: Science advances

Article Title: Retromer promotes the lysosomal turnover of mtDNA.

doi: 10.1126/sciadv.adr6415

Figure Lengend Snippet: Fig. 6. The small GTPase RAB10 promotes mitochondrial fragmentation and mtDNA degradation in lysosomes. (A) Immunostaining of HeLa cells expressing the constitutive active protein RAB10Q68L-GFP labeled with α-VPS35. (B) Manders’ correlation coefficient between RAB10 and VPS35. (C and D) Confocal images of cells ex- pressing WT RAB10-GFP, constitutive active RAB10Q68L-GFP, dominant negative RAB10T23N-GFP, in the steady state, and (D) expressing TWNKK319E-mCherry, labeled with α-TOM20. (E) Quantification of the mitochondrial morphology in RAB10 expressing cells (n = 3, >20 cells per replicate). (F and G) Cells expressing RAB10Q68L-GFP and (G) TWNKK319E-mCherry labeled with α-LAMP1 and α-dsDNA. Arrows depict RAB10-LAMP1-dsDNA foci. (H) Manders’ correlation coefficient between RAB10-GFP and LAMP1 and LAMP1 and dsDNA (n = 3, 10 images per replicate). (I) RAB10-GFP coimmunoprecipitation in the steady state and cells expressing TWNKK319E-mCherry with the lysosomal protein LAMP1. P values were calculated using one-way ANOVA with Tukey correction for multiple comparisons. Scale bars, 10 μm. Data are presented as means ± SEM.

Article Snippet: Antibodies used for immunofluorescence were as follows: polyclonal α- TOM20 (11802- 1- AP; 1:1000) and α- VPS26A (12804- 1- AP; 1:1000) from Proteintech; monoclonal α- dsDNA (ab27156; 1:1000), α- PDHX (ab110333; 1:500), α- ATP5A (ab14748; 1:500), and polyclonal α- RAB5 (1:500) from Abcam; polyclonal α- LAMP1 (#9091; 1:500), α- V5 (#13202; 1:500), and monoclonal α- V5 (#80076; 1:500) from Cell Signaling; monoclonal α- VPS35 (sc- 374372; 1:500) from Santa Cruz.

Techniques: Immunostaining, Expressing, Labeling, Dominant Negative Mutation

Journal: Fish & shellfish immunology

Article Title: Neu1-Deficient Zebrafish Cells Exhibit Reduced Edwardsiella piscicida Infection Due to Altered Lysosomal Exocytosis and Membrane Dynamics.

doi: 10.1016/j.fsi.2025.110273

Figure Lengend Snippet: Fig. 1. Properties of zebrafish primary cells originating from fins. Primary cultured cells were prepared from WT and Neu1-KO zebrafish fins. (A) Morphology of the zebrafish fin cells. (B) Sialidase activity. n = 3. (C) The protein levels of Lamp1 and β-Actin were analyzed by immunoblotting with the cell lysate. (D) Quantitative analysis of the intensities of Lamp1 and β-Actin bands in (C) were carried out and the results are presented as relative Lamp1/β-Actin level to the value in WT cells. n = 4 for each group. (E) Distribution of the Lamp1 protein in cultured fin cells. Lamp1 (red), actin filaments (green), and nuclei (blue) were stained and observed using a fluorescence microscope. The white rectangle shown in the third panel is magnified as the fourth panel. White bar means the scale of 20 μm. White arrows indicate Lamp1 signals in the plasma membrane. Results are shown as means ± standard deviation. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: EGFR, Lamp1, Gapdh, and β-Actin were detected by incubation with rabbit polyclonal antiEGFR antibody (1/1000 dilution, SC-03, Santa Cruz Biotechnoloy, TX, USA), rabbit polyclonal anti-Lamp1 antibody (1/1000, ab24170, Abcam, Cambridge, UK), rabbit polyclonal anti GAPDH antibody (1/ 1000, 60004-1-Ig, Proteintech, IL, USA), and mouse monoclonal antiβ-Actin antibody (1/1000, 66009-1-Ig, Proteintech), respectively, followed by reaction with secondary HRP-anti-mouse or anti-rabbit IgG antibody (1/10,000).

Techniques: Cell Culture, Activity Assay, Western Blot, Staining, Fluorescence, Microscopy, Clinical Proteomics, Membrane, Standard Deviation

Journal: Fish & shellfish immunology

Article Title: Neu1-Deficient Zebrafish Cells Exhibit Reduced Edwardsiella piscicida Infection Due to Altered Lysosomal Exocytosis and Membrane Dynamics.

doi: 10.1016/j.fsi.2025.110273

Figure Lengend Snippet: Fig. 2. Suppression of E. piscicida infection in Neu1-KO cells via the enhanced lysosomal exocytosis. (A) E. piscicida infection in zebrafish primary cells. Results were shown as ratio to colony number in WT cells. n = 10. (B) The protein levels of Lamp1 and Gapdh were analyzed by immunoblotting with the cell lysate with E. piscicida infection. (C) Quantitative analysis of the intensities of Lamp and Gapdh bands in (B) were carried out and the results are presented as relative Lamp1/Gapdh level to the value in WT cells. n = 3 for each group. (D) Distribution of Lamp1 protein in the cultured Neu1-KO cells. Lamp1 (red), actin filament (green), and nucleus (blue) were stained and observed by fluorescence microscopy. White bar means the scale of 20 μm. White arrows indicate the Lamp1 signals at the plasma membrane. (E) E. piscicida infection in Neu1-KO cells with BAPTA-AM pretreatment. Results were shown as ratio to colony number in vehicle (DMSO). n = 10. Results were shown as means ± standard deviation. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: EGFR, Lamp1, Gapdh, and β-Actin were detected by incubation with rabbit polyclonal antiEGFR antibody (1/1000 dilution, SC-03, Santa Cruz Biotechnoloy, TX, USA), rabbit polyclonal anti-Lamp1 antibody (1/1000, ab24170, Abcam, Cambridge, UK), rabbit polyclonal anti GAPDH antibody (1/ 1000, 60004-1-Ig, Proteintech, IL, USA), and mouse monoclonal antiβ-Actin antibody (1/1000, 66009-1-Ig, Proteintech), respectively, followed by reaction with secondary HRP-anti-mouse or anti-rabbit IgG antibody (1/10,000).

Techniques: Infection, Western Blot, Cell Culture, Staining, Fluorescence, Microscopy, Clinical Proteomics, Membrane, Standard Deviation

Journal: Fish & shellfish immunology

Article Title: Neu1-Deficient Zebrafish Cells Exhibit Reduced Edwardsiella piscicida Infection Due to Altered Lysosomal Exocytosis and Membrane Dynamics.

doi: 10.1016/j.fsi.2025.110273

Figure Lengend Snippet: Fig. 1. Properties of zebrafish primary cells originating from fins. Primary cultured cells were prepared from WT and Neu1-KO zebrafish fins. (A) Morphology of the zebrafish fin cells. (B) Sialidase activity. n = 3. (C) The protein levels of Lamp1 and β-Actin were analyzed by immunoblotting with the cell lysate. (D) Quantitative analysis of the intensities of Lamp1 and β-Actin bands in (C) were carried out and the results are presented as relative Lamp1/β-Actin level to the value in WT cells. n = 4 for each group. (E) Distribution of the Lamp1 protein in cultured fin cells. Lamp1 (red), actin filaments (green), and nuclei (blue) were stained and observed using a fluorescence microscope. The white rectangle shown in the third panel is magnified as the fourth panel. White bar means the scale of 20 μm. White arrows indicate Lamp1 signals in the plasma membrane. Results are shown as means ± standard deviation. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: EGFR, Lamp1, Gapdh, and β-Actin were detected by incubation with rabbit polyclonal antiEGFR antibody (1/1000 dilution, SC-03, Santa Cruz Biotechnoloy, TX, USA), rabbit polyclonal anti-Lamp1 antibody (1/1000, ab24170, Abcam, Cambridge, UK), rabbit polyclonal anti GAPDH antibody (1/ 1000, 60004-1-Ig, Proteintech, IL, USA), and mouse monoclonal antiβ-Actin antibody (1/1000, 66009-1-Ig, Proteintech), respectively, followed by reaction with secondary HRP-anti-mouse or anti-rabbit IgG antibody (1/10,000).

Techniques: Cell Culture, Activity Assay, Western Blot, Staining, Fluorescence, Microscopy, Clinical Proteomics, Membrane, Standard Deviation

Journal: Fish & shellfish immunology

Article Title: Neu1-Deficient Zebrafish Cells Exhibit Reduced Edwardsiella piscicida Infection Due to Altered Lysosomal Exocytosis and Membrane Dynamics.

doi: 10.1016/j.fsi.2025.110273

Figure Lengend Snippet: Fig. 2. Suppression of E. piscicida infection in Neu1-KO cells via the enhanced lysosomal exocytosis. (A) E. piscicida infection in zebrafish primary cells. Results were shown as ratio to colony number in WT cells. n = 10. (B) The protein levels of Lamp1 and Gapdh were analyzed by immunoblotting with the cell lysate with E. piscicida infection. (C) Quantitative analysis of the intensities of Lamp and Gapdh bands in (B) were carried out and the results are presented as relative Lamp1/Gapdh level to the value in WT cells. n = 3 for each group. (D) Distribution of Lamp1 protein in the cultured Neu1-KO cells. Lamp1 (red), actin filament (green), and nucleus (blue) were stained and observed by fluorescence microscopy. White bar means the scale of 20 μm. White arrows indicate the Lamp1 signals at the plasma membrane. (E) E. piscicida infection in Neu1-KO cells with BAPTA-AM pretreatment. Results were shown as ratio to colony number in vehicle (DMSO). n = 10. Results were shown as means ± standard deviation. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: EGFR, Lamp1, Gapdh, and β-Actin were detected by incubation with rabbit polyclonal antiEGFR antibody (1/1000 dilution, SC-03, Santa Cruz Biotechnoloy, TX, USA), rabbit polyclonal anti-Lamp1 antibody (1/1000, ab24170, Abcam, Cambridge, UK), rabbit polyclonal anti GAPDH antibody (1/ 1000, 60004-1-Ig, Proteintech, IL, USA), and mouse monoclonal antiβ-Actin antibody (1/1000, 66009-1-Ig, Proteintech), respectively, followed by reaction with secondary HRP-anti-mouse or anti-rabbit IgG antibody (1/10,000).

Techniques: Infection, Western Blot, Cell Culture, Staining, Fluorescence, Microscopy, Clinical Proteomics, Membrane, Standard Deviation

Journal: mBio

Article Title: Coatomer complex I is required for the transport of SARS-CoV-2 progeny virions from the endoplasmic reticulum-Golgi intermediate compartment

doi: 10.1128/mbio.03331-24

Figure Lengend Snippet: Intracellular localization of coatomer complex B2 (COPB2) and ERGIC53 in SARS-CoV-2-infected VeroE6/TMPRSS2 cells. (a–c) VeroE6/TMPRSS2 cells were infected with SARS-CoV-2 at an MOI of 1, fixed at the indicated time points, and subjected to an immunofluorescence assay using antibodies against COPB2 (green in a), ERGIC53 (green in b), LAMP1 (green in c), and SARS-CoV-2 S protein (red). Nuclei were stained with Hoechst 33342 (blue). Box areas shown in merged images are magnified and displayed in the right-hand column. Scale bars: 10 µm. ( D ) Western blot analysis showing the expression levels of SARS-CoV-2 S protein, COPB2, ERGIC53, and β-actin in mock-infected and SARS-CoV-2-infected cells at 16 hpi. ( E ) Immunoelectron microscopy of SARS-CoV-2-infected cells using antibodies against β-COP, followed by secondary antibody conjugated with 6 nm gold beads. (right images) For clarity, gold beads were labeled with red and are shown in the right column. Scale bars: 100 nm. ( F ) Immunoelectron microscopy of SARS-CoV-2-infected cells using antibodies against ERGIC53, followed by secondary antibody conjugated with 6 nm gold beads. For clarity, gold beads were labeled with red and are shown on the right side. Scale bars: 100 nm.

Article Snippet: The commercially available primary antibodies used for immunofluorescence, western blotting, and immuno-electron microscopy were as follows: anti-dsRNA mouse monoclonal antibody J2 (10010200; Scicons; Nordic MUbio, Susteren, Netherlands), anti ERGIC-53 Polyclonal antibody (13364-1-AP, proteintech, IL, USA), anti-β-COP rabbit polyclonal antibody (ab2899; abcam, UK), anti-COPB2 rabbit polyclonal antibody (A304-523A; Bethyl, Montogomery, TX, USA), (ab192924; abcam, UK), anti-SARS-CoV-2-S rabbit polyclonal antibody (NB100-56578; Novus Bililigicals, Centennial, CO, USA), anti-SARS-CoV-2-N rabbit polyclonal antibody (GTX135357; GeneTex, Irvine, CA, USA), anti-SARS-CoV-2-N mouse monoclonal antibody (ZMS1075; Merck, Darmstadt, Germany), anti-LAMP1 rabbit polyclonal antibody (9091T; Cell Signaling Technology, Danvers, MA, USA), anti-Clathrin hevy chain antibody (ab21679, abcam, UK), and anti-β-actin mouse monoclonal antibody (ab8226; abcam, UK).

Techniques: Infection, Immunofluorescence, Staining, Western Blot, Expressing, Immuno-Electron Microscopy, Labeling