anti mouse lamp1 antibody  (Proteintech)

Journal: bioRxiv

Article Title: Spatial Regulation of Lysosomal Vesicle Acidification Along the Axon via mRAVE-Dependent v-ATPase Assembly

doi: 10.64898/2025.12.22.696043

Figure Lengend Snippet: (A) Schematic representation of the v-ATPase complex highlighting the subunits analyzed in this study: V 0 a1 (membrane-embedded V 0 domain) and V 1 B2 (cytosolic V 1 domain). Fluorescent proteins (FP) were fused to the C-terminus of the V 0 a1 subunit and the N-terminus of the V 1 B2 subunit. (B) Immunofluorescence microscopy of HeLa cells stably transfected with V 0 a1–eGFP or AcGFP–V 1 B2 constructs. Single-channel images are shown in grayscale, with DAPI (nuclei) shown in blue. (C) Quantification of the peripheral/perinuclear mean intensity of V 0 a1–GFP and AcGFP–V 1 B2 from experiments such as that shown in panel B (n=29-32 cells from three independent experiments). (D,F) Immunofluorescence microscopy showing co-localization of stably expressed V 0 a1–eGFP (green) (D) or AcGFP–V 1 B2 (green) (F) with endogenous LAMP1 (magenta). Nuclei were stained with DAPI (blue). Magnified views of the boxed areas in the peripheral and perinuclear regions are shown at left and right, respectively. (E,G) Quantification of the co-localization between LAMP1 and V 0 a1–eGFP (E) or AcGFP–V 1 B2 (G) in total, perinuclear, and peripheral regions, expressed as Pearson’s correlation coefficient (Pearson’s r), from experiments such as those shown in panels D and F ( n =22 cells from three independent experiments). (H) Live-cell images of HeLa cells showing the co-localization of transiently co-expressed V 0 a1–mScarlet3 (magenta) and AcGFP–V 1 B2 (green). Magnified views of the boxed areas in the peripheral and perinuclear regions are shown at left and right, respectively . (I) Quantification of Pearson’s correlation coefficients between V 0 a1–mScarlet3 and AcGFP–V 1 B2 from experiments such as that shown in panel H ( n =23 cells from three independent experiments). All quantitative data are represented as the mean ± SD. Statistical significance was assessed using the Welch’s t-test for panel C, and Friedman test with Dunn’s multiple comparisons test for panels E, G, and I. Actual P values are indicated in the figure. Scale bars: 10μm.

Article Snippet: Primary antibodies: mouse anti-DMXL2 (Proteintech, Cat. 66891-2-Ig, 1:1,000 for IB), rabbit anti-ATP6V0A1 (Novus Biologicals, Cat. NBP1-89342, 1:1,000 for IB), rabbit anti-ATP6V1B2 (Abcam, Cat. ab73404, 1:1,000 for IB), mouse anti-human LAMP1 (DSHB, Cat. H4A3, 1:1,000 for IF), mouse anti-rat LAMP1 (6H2, generated in-house, 1:1 culture supernatant for IF), chicken anti-GFP (Invitrogen, Cat. A10262, 1:1,000 for IF), rat anti-mCherry (Invitrogen, Cat. M11217, 1:500 for IF), goat anti-ankyrin G (Santa Cruz Biotechnology, Cat. sc-31778, 1:100 for IF), rabbit anti-LC3 (Cell Signaling, Cat. 3868, 1:200 for IF) and anti-pan-neurofascin (extracellular) antibody (A12/18) (Antibodies Inc, Cat. 74-172) labeled with the mix-n-Stain Neurofascin CF647 Antibody Labeling Kit (Biotium, Cat. 92238), according to the manufacturer’s protocol (to label the axon initial segment for live-cell imaging).

Techniques: Membrane, Immunofluorescence, Microscopy, Stable Transfection, Transfection, Construct, Staining

Journal: bioRxiv

Article Title: Spatial Regulation of Lysosomal Vesicle Acidification Along the Axon via mRAVE-Dependent v-ATPase Assembly

doi: 10.64898/2025.12.22.696043

Figure Lengend Snippet: (A,C) Immunofluorescence microscopy showing co-localization of stably expressed V 0 a1–eGFP (green) (A) or AcGFP–V 1 B2 (green) (C) with endogenous LAMP1 (magenta) in U2OS cells. Nuclei were stained with DAPI (blue). Magnified views of the boxed areas in the peripheral and perinuclear regions are shown at left and right, respectively. (B,D) Quantification of the co-localization between LAMP1 and V 0 a1–eGFP (B) or AcGFP–V 1 B2 (D) in total, perinuclear, and peripheral regions, expressed as Pearson’s correlation coefficients (Pearson’s r), from experiments such as those shown in panels A and C ( n =23-25 cells from three independent experiments). (E) Single frame live-cell images of HeLa cells showing the co-localization of stably expressed V 0 a1–eGFP (green) with LysoTracker (magenta). Magnified views of the boxed areas in the peripheral and perinuclear regions are shown at left and right, respectively . (F) Quantification of the co-localization between LysoTracker and V 0 a1–eGFP in total, perinuclear, and peripheral regions, expressed as Pearson’s r, from experiments such as those shown in panel E ( n =18-19 cells from two independent experiments). All quantitative data are represented as the mean ± SD. Statistical significance was assessed using the Friedman test with Dunn’s multiple comparisons test. Actual P values are indicated in the figure. Scale bars: 10 μm.

Article Snippet: Primary antibodies: mouse anti-DMXL2 (Proteintech, Cat. 66891-2-Ig, 1:1,000 for IB), rabbit anti-ATP6V0A1 (Novus Biologicals, Cat. NBP1-89342, 1:1,000 for IB), rabbit anti-ATP6V1B2 (Abcam, Cat. ab73404, 1:1,000 for IB), mouse anti-human LAMP1 (DSHB, Cat. H4A3, 1:1,000 for IF), mouse anti-rat LAMP1 (6H2, generated in-house, 1:1 culture supernatant for IF), chicken anti-GFP (Invitrogen, Cat. A10262, 1:1,000 for IF), rat anti-mCherry (Invitrogen, Cat. M11217, 1:500 for IF), goat anti-ankyrin G (Santa Cruz Biotechnology, Cat. sc-31778, 1:100 for IF), rabbit anti-LC3 (Cell Signaling, Cat. 3868, 1:200 for IF) and anti-pan-neurofascin (extracellular) antibody (A12/18) (Antibodies Inc, Cat. 74-172) labeled with the mix-n-Stain Neurofascin CF647 Antibody Labeling Kit (Biotium, Cat. 92238), according to the manufacturer’s protocol (to label the axon initial segment for live-cell imaging).

Techniques: Immunofluorescence, Microscopy, Stable Transfection, Staining

Journal: bioRxiv

Article Title: Spatial Regulation of Lysosomal Vesicle Acidification Along the Axon via mRAVE-Dependent v-ATPase Assembly

doi: 10.64898/2025.12.22.696043

Figure Lengend Snippet: (A) Schematic representation of the isolation, transfection, and processing of embryonic rat hippocampal neurons. (B) Immunofluorescence microscopy of DIV7 rat hippocampal neurons transiently transfected with a plasmid encoding V 0 a1–eGFP and immunostained for the endogenous lysosomal membrane protein LAMP1 (magenta) and axon initial segment (AIS) protein ankyrin G (AnkG) (blue). Magnified views of the boxed 30-μm axonal segments are shown at right. (C) Quantification of V 0 a1–eGFP, LAMP1, and V 0 a1–eGFP-positive LAMP1 vesicles per 30 μm axon length from experiments such as that shown in panel B ( n =18 neurons from ≥4 cultures prepared from two rats). (D) Immunofluorescence microscopy of DIV7 rat hippocampal neurons transfected with a plasmid encoding AcGFP–V 1 B2 and immunostained for endogenous LAMP1 (magenta) and ankyrin G (blue). Magnified views of the boxed 30-μm axonal segments are shown at right. (E) Quantification of AcGFP–V 1 B2, LAMP1, and AcGFP–V 1 B2-positive LAMP1 vesicles per 30 μm axon length from experiments such as that shown in panel D ( n =16 neurons from ≥4 cultures prepared from two rats). (F) Immunofluorescence microscopy of DIV7 rat hippocampal neurons co-transfected with plasmids encoding V 0 a1–eGFP and mCherry–V 1 B2. Magnified views of the boxed 30-μm axonal segments are shown at right. (G) Quantification of V 0 a1–eGFP, mCherry–V 1 B2, and mCherry–V 1 B2-positive V 0 a1–eGFP vesicles per 30 μm axon length from experiments such as that shown in panel F ( n =17 neurons from ≥4 cultures prepared from two rats). All the axonal segments analyzed correspond to the mid-axon and are located approximately 40-200 μm from the soma. All quantitative data are represented as the mean ± SD. Statistical significance was assessed using the Friedman test with Dunn’s multiple comparisons test. Actual P values are indicated in the figure. Scale bars: 20 μm.

Article Snippet: Primary antibodies: mouse anti-DMXL2 (Proteintech, Cat. 66891-2-Ig, 1:1,000 for IB), rabbit anti-ATP6V0A1 (Novus Biologicals, Cat. NBP1-89342, 1:1,000 for IB), rabbit anti-ATP6V1B2 (Abcam, Cat. ab73404, 1:1,000 for IB), mouse anti-human LAMP1 (DSHB, Cat. H4A3, 1:1,000 for IF), mouse anti-rat LAMP1 (6H2, generated in-house, 1:1 culture supernatant for IF), chicken anti-GFP (Invitrogen, Cat. A10262, 1:1,000 for IF), rat anti-mCherry (Invitrogen, Cat. M11217, 1:500 for IF), goat anti-ankyrin G (Santa Cruz Biotechnology, Cat. sc-31778, 1:100 for IF), rabbit anti-LC3 (Cell Signaling, Cat. 3868, 1:200 for IF) and anti-pan-neurofascin (extracellular) antibody (A12/18) (Antibodies Inc, Cat. 74-172) labeled with the mix-n-Stain Neurofascin CF647 Antibody Labeling Kit (Biotium, Cat. 92238), according to the manufacturer’s protocol (to label the axon initial segment for live-cell imaging).

Techniques: Isolation, Transfection, Immunofluorescence, Microscopy, Plasmid Preparation, Membrane

Journal: Nature Communications

Article Title: Ubiquitination by HRD1 is essential for TLR3 trafficking and its innate immune signaling

doi: 10.1038/s41467-025-67219-0

Figure Lengend Snippet: a – f Confocal microscopic images of TLR3 co-stained with DAPI and ER marker KDEL ( a ), trans-Golgi network marker TGN38 ( b ) and lysosome marker LAMP1 ( c ) in WT and Hrd1 −/− RAW 264.7 cells with or without poly(I:C) (50 µg/ml) treatment. Quantitation of the fraction of TLR3 in the ER ( d , from left to right, n = 60, 58, 60, 60, 60, 60), trans-Golgi network ( e , n = 60 for all groups), and lysosomes ( f , n = 60 for all groups) in these macrophages were shown. Mander’s overlap coefficient is used for measurement of colocalization. g – j Confocal microscopic images of TLR3-Flag co-stained with DAPI and ER marker Calnexin ( g ), Golgi marker GM130 ( h ), early endosome marker Rab5 ( i ) and late endosome marker Rab7 ( j ) in WT and Hrd1 −/− HEK293T cells transfected with TLR3-Flag plasmid and with or without poly(I:C) treatment. k – m Immunoblot analysis of full-length (FL) and cleaved TLR3 (CL) in WT and Hrd1 −/− HEK293T cells transfected with TLR3-Flag plasmid and treated with 50 μg/ml poly(I:C) for 12 h ( k ), LS-102 (5 μM) for 8 h ( l ), and with Endo H or PNGase F ( m ). Arrow, Endo H-sensitive; Red box, Endo H-resistant. n Immunoblot analysis of indicated proteins following the isolation of the ER, Golgi and endosomes from WT and Hrd1 −/− HEK293T cells transfected with TLR3-Flag for 24 h, followed by 50 µg/ml poly(I:C) stimulation for 1 h. Relative quantitation of indicated TLR3 bands shown below the blot. o – q Immunoblot analysis of indicated proteins following immunoprecipitation of TLR3-Flag in the lysates of HEK293T cells transfected with TLR3-Flag, HRD1-myc (WT, C2A, and ΔRING), and HRS-V5 ( o ), or TGS101-V5 ( p ), or VPS36-V5 ( q ). C2A, HRD1-dead variant; ΔRING, RING domain-deleted truncate. Quantitation of the protein level is shown below the blot. All confocal image and blot data were representative of at least three independent repeats ( a – q ). Values represent mean ± SEM, by unpaired, two-tailed, Student’s t -test ( d – f ). Source data are provided as a Source Data file.

Article Snippet: Primary antibodies for immunofluorescent staining included: TLR3 (Proteintech 17766-1-AP, 1: 200), Calnexin (Proteintech 10427-2-AP, 1: 200), GM130 (Proteintech 11308-1-AP, 1:1000), Rab5 (Cell Signaling C8B1; 1: 200), Rab7 (Cell Signaling D95F2; 1: 200), KDEL (Novus Biologicals NBP1-97469, 1:200), TGN38 (Santa Cruz sc-166594,1:200), LAMP1 (Proteintech 65050-1-Ig, 1:1000), Flag (Sigma F1804, 1: 200).

Techniques: Staining, Marker, Quantitation Assay, Transfection, Plasmid Preparation, Western Blot, Isolation, Immunoprecipitation, Variant Assay, Two Tailed Test

Journal: Autophagy Reports

Article Title: Autophagy cargo profiles in skeletal muscle during starvation and exercise

doi: 10.1080/27694127.2025.2593060

Figure Lengend Snippet: Mitophagy and ribophagy in skeletal muscles under basal, starvation and exercise conditions. (A) (B) Mitophagy levels in basal, starvation and exercise conditions assessed in the muscles of Mito-QC mice. The numbers of red-only dots reflect the mitophagy flux and were quantified in (B). Data points are individual mice ( n = 4). * p < 0.05 using two sides, unpaired Student’s t-test when comparing with basal condition. & p < 0.05 using two sides, unpaired Student’s t-test when comparing with starvation condition. Data are mean ± s.e.m. Scale bar, 20 μm. (C) (D) Ribophagy levels in basal, starvation and exercise conditions assessed by examining the colocalization of ribosome protein S6 (RPS6) with lysosome protein LAMP1 in the muscles of wild-type mice. The numbers of colocalization dots per muscle fiber were quantified in (D). Data points are individual mice ( n = 3), and 40 to 60 fibers were quantified for each mouse. * p < 0.05 using two sides, unpaired Student’s t-test when comparing with basal condition. & p < 0.05 using two sides, unpaired Student’s t-test when comparing with exercise condition. Data are mean ± s.e.m. Scale bar, 10 μm. Arrowheads denote the colocalization dots.

Article Snippet: Anti-COXIV (11242–1-AP), anti-GAPDH (60004–1-Ig), anti-GM130 (11308–1-AP), anti-Cathepsin D (21327–1-AP), anti-ATP1A2 (16836–1-AP), anti-mouse LAMP1 (65050–1-Ig), anti-FAM134B (21537–1-AP) and anti-TEX264 (25858–1-AP) antibodies were from Proteintech.

Techniques: Muscles

Journal: Frontiers in Molecular Biosciences

Article Title: Loss of ARF5 impairs recovery after lysosomal damage

doi: 10.3389/fmolb.2025.1699266

Figure Lengend Snippet: ARF GTPases are recruited to damaged lysosomes. (A) HeLa cells were transfected with HA-tagged ARF1, ARF5, or ARF6 and treated the next day with 0.5 mM LLOME for 1 h or 3 h. At each time point, cells were fixed and stained for HA (cyan) and endogenous LAMP1 (magenta) and imaged with super-resolution confocal microscopy (Nikon NSPARC system). Scale bar indicates 10 µm for original size images and 1 µm for zoomed images. (B) Quantification of data in (A) n = 15 cells. Error bars represent mean +SD. Data were analyzed with one-way ANOVA. *p < 0.05 (C) (left) HEK293 cells stably expressing 3XHA-tagged TMEM192 were transfected with GFP-tagged ARF1, ARF5, or ARF6 constructs. Untreated and LLOME-treated (3 h) cells were subjected to lysosome immunoprecipitation using HA-conjugated magnetic beads (LysoIP). Total cell lysates and immunoprecipitates were immunoblotted for GFP to detect bound ARFs (left). Lysates and immunoprecipitates were also immunoblotted for endogenous LAMP1, Golgin-97, or EEA1 (right). (D) Quantification of data in (C) . Bars indicate mean +SD of ≥2 experimental replicates. Statistical analysis was performed using unpaired t-test. *p < 0.05. Statistics represent comparison of untreated HEK293 cells with 3-h LLOME (1 mM) treated cells quantified by densitometry. (E) HEK293 cells were treated with LLOME or not and subjected to the LysoIP protocol. Lysates were immunoblotted and stained for endogenous ARF5 and HA. (F) Quantification of data in (E) . Bars indicate mean +SD of three experimental replicates. *p < 0.05. Statistics represent comparison of untreated HEK293 cells with 20-min and 3-h LLOME-treated cells quantified by densitometry.

Article Snippet: The antibodies we used were: rabbit antibody to LAMP1 (D2D11) (Cell Signaling Technology, Inc.), Cat No. 9091S, IF (1:400) WB (1:1200); mouse antibody to LAMP1 (1D4B) from Developmental Studies Hybridoma Bank; mouse antibody to GFP (Proteintech) Cat No.:66002-1-Ig WB (1:100,000); mouse antibody to HA (16B12) (Biolegend) WB (1:4000); rabbit antibody to mCherry (Sigma-Aldrich) Cat No.SAB2702295-100UL WB (1:10,000); rabbit antibody to OSBP (Sigma) Cat no. HPA039227 WB (1:1100) IF (1:100); rabbit antibody to ORP9 from Dr. Neale Ridgway, Dalhousie University, IF (1:1000); mouse antibody to Golgin97 (Molecular probes) Cat No. CDF4 A-21270 WB (1:500); rabbit antibody to ARF5 (Novus Biologicals) Cat No. NBP1-31005 WB (1:2500); sheep antibody to TGN46 (Serotec, Oxford United Kingdom); mouse antibody to Gal-3 (B-2) (Santa Cruz Biotechnology, Inc.) Cat No. sc-25279 IF (1:100); IRDye 800CW donkey anti-mouse secondary antibody (Li-COR) 926-32212 WB (1:10,000); IRDye 680RD goat anti-rabbit secondary antibody (Li-COR) 926-68071 WB (1:10,000); Alexa Fluor 488 donkey anti-mouse (ThermoFisher Scientific (Rockford, IL) IF (1:100); Alexa Fluor 488 donkey anti-rabbit (ThermoFisher Scientific (Rockford, IL) IF (1:100); Alexa Fluor 568 donkey anti-mouse (ThermoFisher Scientific (Rockford, IL) IF (1:100); Alexa Fluor 568 donkey anti-rabbit (ThermoFisher Scientific (Rockford, IL) IF (1:100).

Techniques: Transfection, Staining, Confocal Microscopy, Stable Transfection, Expressing, Construct, Immunoprecipitation, Magnetic Beads, Comparison

Journal: Frontiers in Molecular Biosciences

Article Title: Loss of ARF5 impairs recovery after lysosomal damage

doi: 10.3389/fmolb.2025.1699266

Figure Lengend Snippet: Lipid transfer proteins OSBP and ORP9 are recruited to damaged lysosomes in an ARF-independent manner. (A) HeLa cells were transfected with empty vector (shCTRL) or pLKO containing ARF1 or ARF5 shRNAs. Cells were treated or not with LLOME (0.5 mM) for 35 min, fixed and stained for endogenous OSBP (green), LAMP1 (red), and TGN46 (magenta), and imaged with confocal microscopy. Scale bars indicate 10 µm. (B) HEK293 cells stably expressing tagged TMEM192 were transfected with empty pLKO vector (shCTRL) or pLKO containing ARF1 or ARF5 shRNAs. Untreated and LLOME (1 mM)-treated cells were subjected to the LysoIP protocol. Precipitated lysosomes were immunoblotted for endogenous OSBP. (C) HeLa cells were transfected with empty vector (shCTRL), ARF5, or ARF6 shRNAs. Cells were treated or not with LLOME (0.5 mM) for 20 min, fixed and stained for ORP9 (green), LAMP1 (red), and F-actin (blue), and imaged with confocal microscopy. Scale bars indicate 10 µm. (D) Quantification of data in (C) n = 28 cells. Error bars represent mean +SD. Data were analyzed with two-way ANOVA with Dunnett’s multiple comparisons test with a single pooled variance. *p < 0.05.

Article Snippet: The antibodies we used were: rabbit antibody to LAMP1 (D2D11) (Cell Signaling Technology, Inc.), Cat No. 9091S, IF (1:400) WB (1:1200); mouse antibody to LAMP1 (1D4B) from Developmental Studies Hybridoma Bank; mouse antibody to GFP (Proteintech) Cat No.:66002-1-Ig WB (1:100,000); mouse antibody to HA (16B12) (Biolegend) WB (1:4000); rabbit antibody to mCherry (Sigma-Aldrich) Cat No.SAB2702295-100UL WB (1:10,000); rabbit antibody to OSBP (Sigma) Cat no. HPA039227 WB (1:1100) IF (1:100); rabbit antibody to ORP9 from Dr. Neale Ridgway, Dalhousie University, IF (1:1000); mouse antibody to Golgin97 (Molecular probes) Cat No. CDF4 A-21270 WB (1:500); rabbit antibody to ARF5 (Novus Biologicals) Cat No. NBP1-31005 WB (1:2500); sheep antibody to TGN46 (Serotec, Oxford United Kingdom); mouse antibody to Gal-3 (B-2) (Santa Cruz Biotechnology, Inc.) Cat No. sc-25279 IF (1:100); IRDye 800CW donkey anti-mouse secondary antibody (Li-COR) 926-32212 WB (1:10,000); IRDye 680RD goat anti-rabbit secondary antibody (Li-COR) 926-68071 WB (1:10,000); Alexa Fluor 488 donkey anti-mouse (ThermoFisher Scientific (Rockford, IL) IF (1:100); Alexa Fluor 488 donkey anti-rabbit (ThermoFisher Scientific (Rockford, IL) IF (1:100); Alexa Fluor 568 donkey anti-mouse (ThermoFisher Scientific (Rockford, IL) IF (1:100); Alexa Fluor 568 donkey anti-rabbit (ThermoFisher Scientific (Rockford, IL) IF (1:100).

Techniques: Transfection, Plasmid Preparation, Staining, Confocal Microscopy, Stable Transfection, Expressing

Journal: Frontiers in Molecular Biosciences

Article Title: Loss of ARF5 impairs recovery after lysosomal damage

doi: 10.3389/fmolb.2025.1699266

Figure Lengend Snippet: ARF5-depleted cells do not recover from lysosomal damage. (A) Schematic of Gal3 recovery assay. (B) HeLa cells were transfected with empty vector control (shCTRL), ARF5, or ARF6 shRNAs. Cells were treated or not with LLOME (0.5 mM) which was rinsed out after 20 min of incubation. Cells were allowed to recover for either 1 h or 8 h, after which they were fixed and stained for endogenous Gal3 (cyan), LAMP1 (magenta), and F-actin (yellow) and imaged with confocal microscopy. Scale bar indicates 10 µm. (C) Quantification of data in (B) n = 40–66 cells from two independent experiments. Error bars represent mean +SD. Data were analyzed with nonparametric Kruskal–Wallis one-way ANOVA test with Dunn’s correction. *p < 0.05 (D) HeLa cells were transfected with empty vector (shCTRL), ARF5, or ARF6 shRNAs and plated in black-walled 96-well plates. Cells were then treated with LLOME overnight or after 20 min of incubation, rinsed and allowed to recover (LLOME + recovery). Cells were incubated overnight, fixed the next day, and stained for actin (Alexa-488 phalloidin). Fluorescence was analyzed using a BioTek imaging microplate reader. y-axis represents Alexa-488 nm fluorescence [AU]. Data were analyzed with two-way ANOVA with Dunnett’s multiple comparisons test with a single pooled variance. *p < 0.05.

Article Snippet: The antibodies we used were: rabbit antibody to LAMP1 (D2D11) (Cell Signaling Technology, Inc.), Cat No. 9091S, IF (1:400) WB (1:1200); mouse antibody to LAMP1 (1D4B) from Developmental Studies Hybridoma Bank; mouse antibody to GFP (Proteintech) Cat No.:66002-1-Ig WB (1:100,000); mouse antibody to HA (16B12) (Biolegend) WB (1:4000); rabbit antibody to mCherry (Sigma-Aldrich) Cat No.SAB2702295-100UL WB (1:10,000); rabbit antibody to OSBP (Sigma) Cat no. HPA039227 WB (1:1100) IF (1:100); rabbit antibody to ORP9 from Dr. Neale Ridgway, Dalhousie University, IF (1:1000); mouse antibody to Golgin97 (Molecular probes) Cat No. CDF4 A-21270 WB (1:500); rabbit antibody to ARF5 (Novus Biologicals) Cat No. NBP1-31005 WB (1:2500); sheep antibody to TGN46 (Serotec, Oxford United Kingdom); mouse antibody to Gal-3 (B-2) (Santa Cruz Biotechnology, Inc.) Cat No. sc-25279 IF (1:100); IRDye 800CW donkey anti-mouse secondary antibody (Li-COR) 926-32212 WB (1:10,000); IRDye 680RD goat anti-rabbit secondary antibody (Li-COR) 926-68071 WB (1:10,000); Alexa Fluor 488 donkey anti-mouse (ThermoFisher Scientific (Rockford, IL) IF (1:100); Alexa Fluor 488 donkey anti-rabbit (ThermoFisher Scientific (Rockford, IL) IF (1:100); Alexa Fluor 568 donkey anti-mouse (ThermoFisher Scientific (Rockford, IL) IF (1:100); Alexa Fluor 568 donkey anti-rabbit (ThermoFisher Scientific (Rockford, IL) IF (1:100).

Techniques: Transfection, Plasmid Preparation, Control, Incubation, Staining, Confocal Microscopy, Fluorescence, Imaging