Anti-LAMP2 antibody [GL2A7] - Lysosome Marker

• WB

AbReview29449****

Western blot - Anti-LAMP2 antibody [GL2A7] - Lysosome Marker (AB13524)

Blocked with 5% milk for 1 hour at RT.

Incubated with primary antibody in 5% BSA/TBST for 16 hours at 4°C.

All lanes:

Western blot - Anti-LAMP2 antibody [GL2A7] - Lysosome Marker (ab13524) at 1/1000 dilution

All lanes:

HEK293 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-rat HRP at 1/5000 dilution

Predicted band size: 45 kDa

Observed band size: 105 kDa,28 kDa,70 kDa

true

Exposure time: 3min

This image is courtesy of an anonymous Abreview.

• WB

Supplier Data

Western blot - Anti-LAMP2 antibody [GL2A7] - Lysosome Marker (AB13524)

Block : 5% milk + TBST for 1 hour at RT.

All lanes:

Western blot - Anti-LAMP2 antibody [GL2A7] - Lysosome Marker (ab13524) at 1/500 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate at 20 µg

Lane 2:

NIH/3T3 (Mouse embryo fibroblast cell line) lysate at 10 µg

Secondary

All lanes:

HRP Goat Anti-Rat, 1 hour at RT at 1/100 dilution

Predicted band size: 45 kDa

false

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-LAMP2 antibody [GL2A7] - Lysosome Marker (AB13524)

Immunocytochemistry-immunofluorescence using Anti-LAMP2 antibody [GL2A7], ab13524. Publication image from Zhao, T. et al., 2015, Nat Commun, 26437053. Legend direct from paper.

Buffering the pH of endocytic organelles affects their membrane protein dynamics.HeLa cells were treated with 500 µg ml−1 dextran-TMR or 1,000 µg ml−1 UPS6.2-Cy5 or UPS4.4-Cy5 for 5 min for cell uptake. Then they were fixed after 15 min (a), 1 h (b) and 2 h (c). Immunofluorescence (IF) images show the localization of UPS nanoparticles in early endosomes (Rab5) or lysosomes (LAMP2). Scale bar, 10 and 5 µm (inset). Imaris software was used to analyse co-localization of z-stacked confocal images. The fraction of UPS/dextran co-localized with Rab5 (d) and LAMP2 (e) and the fraction of Rab5 co-localized with LAMP2 (f) were calculated from thresholded Mander's coefficient (see Supplementary Methods), n=10,α=0.05, ****P<0.0001. Two-way analysis of variance and Sidak's multiple comparison tests were performed to assess the statistical significance. The dashed line in (f) represents the basal level of Rab 5 and LAMP2 co-localization in HeLa cells without any treatment.

• Flow Cyt

CiteAb

Flow Cytometry - Anti-LAMP2 antibody [GL2A7] - Lysosome Marker (AB13524)

Flow cytometry/Cell sorting using Anti-LAMP2 antibody [GL2A7], ab13524. Publication image from Weihl, C. C. et al., 2017, Nat Commun, 28589926. Legend direct from paper.

TFEB overexpression in macrophages induces the autophagy markers LC3 and p62 and restores their co-localization in atherosclerotic aortic roots.(a,b) Representative immunofluorescence images of atherosclerotic aortic roots (2 months' western diet) from control and mφTFEB-TG mice (ApoE-null background) stained with antibodies against TFEB (a), TFEB and MOMA-2 (b; scale bar, 50 µm). (c) Quantification of the average TFEB intensity and co-localization with nuclear marker DAPI (n=4-5 mice per group). (d) Representative immunofluorescence images of atherosclerotic aortic roots from control and mφTFEB-TG mice stained with p62 and LC3 (scale bar, 50 µm). (e) Quantification of the p62 and LC3 average intensity from control and mφTFEB-TG-stained roots (n=13–14 mice per group). (f) Representative pseudocolour image of these p62/LC3 images (green represents co-localization) and graph depicting the increased p62/LC3 correlation seen in a representative mφTFEB-TG as compared to a control lesion (scale bar, 50 µm). (g) Quantification of the p62/LC3 co-localization from control and mφTFEB-TG-stained roots shown (n=13–14 mice per group). (h,i) FACS analysis of aortic macrophages isolated from atherosclerotic aortas of Control or mφTFEB-TG mice (western diet-fed ApoE-KO background, n=3–4 pooled aortas) and stained for either (h) p62 and LC3, or (i) Lamp2 and LC3 antibodies (per cent of macrophages expressing each marker is shown below plots). For all graphs, data are presented as mean±s.e.m. *P<0.05, ***P<0.001, two-tailed unpaired t-test.

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-LAMP2 antibody [GL2A7] - Lysosome Marker (AB13524)

LAMP2 Immunocytochemistry-immunofluorescence using Anti-LAMP2 antibody [GL2A7] ab13524. Publication image and figure legend from Mizunoe, Y., Kobayashi, M., et al., 2020, Sci Rep, Pubmed 31959889.

Localization of PLIN1 in CTSB-OE 3T3L1 adipocytes. (A and B) Immunofluorescence analysis was performed with anti-LAMP2 antibodies in CTSB-mCherry-OE 3T3L1 adipocytes. Images of CTSB-mCherry (red) and LAMP2 (green) (A), and histogram of fluorescence intensity (B) are shown. Representative images of individual experiments are shown. Scale bar represents 10 μm. Asterisk indicates LD. (C) CTSB-OE 3T3L1 adipocytes were treated with 10 μm CA074ME for 24 h. Immunofluorescence analysis was performed with anti-PLIN1 and LAMP2 antibodies. PLIN1 (green) and LAMP2 (red) are shown. Representative images of individual experiments are shown. Scale bar represents 5 μm. Arrows indicate the contact site between PLIN1 and LAMP2. (D) CTSB-mCherry-OE 3T3L1 adipocytes were treated with 10 μm CA074ME for 24 h. Immunofluorescence analysis was performed with an anti-PLIN1 antibody. PLIN1 (green) and CTSB-mCherry (red) are shown. Representative images of individual experiments are shown. Scale bar represents 5 μm. Arrows indicate the contact site between PLIN1 and CTSB-mCherry.

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-LAMP2 antibody [GL2A7] - Lysosome Marker (AB13524)

LAMP2 immunocytochemistry/ immunofluorescence using Anti-LAMP2 antibody [GL2A7] ab13524. Publication image and figure legend from Peelaerts, W., Bergkvist, L., et al., 2020, Free Neuropathol, Pubmed 35224554.

Absence of inflammatory or autophagy deficits 13-weeks post PFF surgery.a) Representative images of Iba-1 positive staining in the ipsilateral AON, scale bar = 50 μm. b) Quantification of microglial hydraulic radius as a measure of microglial ramification and activation (n = 5-6/group, SEM), shows no microglial activation following PFF injection. c) Representative images of LAMP2 positive staining (green) in the ipsilateral AON. DAPI (blue) was used to visualize the cell nuclei. Scale bar = 10 μm d) Fluorescent LAMP2 quantification after adding adaptive triangle thresholding in the AON (n = 6, SEM)

• WB

CiteAb

Western blot - Anti-LAMP2 antibody [GL2A7] - Lysosome Marker (AB13524)

Western Blotting using Anti-LAMP2 antibody [GL2A7], ab13524. Publication image from Iershov, A. et al., 2019, Nat Commun, 30952952. Legend direct from paper.

PPARα activation by fenofibrate restores lipid catabolism in Vps15-LKO mice. a Relative transcript levels of metabolic enzymes in ketogenesis, FAO, fatty acid transport and peroxisome biogenesis in livers of fed Vps15f/f and AlbCre+;Vps15f/f mice that were treated for two weeks with fenofibrate. Data are means ± SEM (Vps15f/f (n = 6 and n = 4 for chow and FENO group), AlbCre+;Vps15f/f (n = 5 and n = 4 for chow and FENO group), P < 0.05 * : vs Vps15f/f, # : vs chow, two-tailed, unpaired Student’s t test). b Immunoblot analysis of total protein liver extracts of mice treated as in a using indicated antibodies. Densitometric analyses of protein levels normalised to Pras40 levels presented as folds over Vps15f/f-chow condition. Data are means ± SEM (n = 4 for Vps15f/f chow and FENO group, n = 3 for AlbCre+;Vps15f/f chow and n = 4 for AlbCre+;Vps15f/f FENO group, P < 0.05 * : vs Vps15f/f, # : vs chow, two-tailed, unpaired Student’s t test). c Representative images of immunofluorescent analyses showing nuclear localization of PPARα, NCoR1 and Hdac3 in liver tissue of Vps15f/f and AlbCre+;Vps15f/f mice treated as in a. Secondary anti-mouse or anti-rabbit IgG Alexa Fluor 568 antibody were used for detection. Scale bar : 40 µm. d Relative levels of Acetyl-CoA and Acyl-carnitine metabolites measured by mass spectrometry in liver tissue of mice treated as in a. Data are means ± SEM (Vps15f/f (n = 4 for chow and FENO group), AlbCre+;Vps15f/f (n = 3 and n = 4 for chow and FENO group, P < 0.05 * : vs Vps15f/f, # : vs chow, two-tailed, unpaired Student’s t test). e Immunoblot analyses in total protein liver extracts of 6-week old Vps15f/f and AlbCre+;Vps15f/f mice. Mice were either fed or fasted for 24 h. Four hours prior the sacrifice fasted mice were injected with leupeptin (40 mg/kg) or vehicle. The total protein extracts were immunoblotted with indicated antibodies. f Immunoblot analysis of cytosolic protein fractions of primary hepatocytes that were grown for 72 h in fasting media and treated for 24 h before collection with increasing doses of BafA1. Densitometric analyses of protein levels normalised to Tubulin levels presented as folds over vehicle-treated cells. Data are means ± SEM (n = 4 with 100 nM Bafilomycin A1 treatment, P < 0.05 * : vs vehicle, two-tailed, unpaired Student’s t test). g Relative transcript levels (left panel) of indicated genes in primary hepatocytes incubated in control or fasting media (72 h) treated with or without BafA1 for 24 h before collection. Data are means ± SEM (n = 3, P < 0.05 * : vs control media, # : vs fasting media, two-tailed, unpaired Student’s t test). The immunoblot (right panel) served as control of autophagic activity

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• WB

CiteAb

Western blot - Anti-LAMP2 antibody [GL2A7] - Lysosome Marker (AB13524)

LAMP2 western blotting using Anti-LAMP2 antibody [GL2A7] ab13524. Publication image and figure legend from Li, Z., Liu, S., et al., 2019,Stem Cell Res Ther, Pubmed 31775910.

High concentration of silicate did not affect lysosome function of BMSCs. a, b Lyso-tracker assay, which could lock acid lysosomes, was applied to evaluate the function of lysosomes, and the Baf A1 and CQ treatment groups were set as the positive control groups (n = 3, 30 random cells per sample). The data indicated that the fluorescence intensity in the silicate (3 mM)-treated groups was similar to that of the CQ-treated group, and the intensity increased significantly compared with that of the control group. c, d Western blot was performed to characterize the expression of LAMP1 and LAMP2 in BMSCs after exposure to silicate for 24 h (n = 4). Data analysis showed that LAMP1 and LAMP2 increased significantly in the high-concentration silicate group. (*p < 0.05, **p < 0.01, ***p < 0.001)

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