antibody against lamp1  (Boster Bio)

Journal: bioRxiv

Article Title: Mechanoresilience of lysosomes conferred by TMEM63A

doi: 10.64898/2025.12.18.695245

Figure Lengend Snippet: A) LysoFlipper FLIM in WT RAW264.7 macrophages incubated with 1 mM LLOMe for 5 or 15 min, or 250 µM GPN for 15 min. Each dot represents one field of 5-10 cells. N = 3. B) qPCR for TMEM63A, B, and C in RAW264.7, BMDM, or MEF. n = 3. C-D) TMEM63A-OFP (magenta) and LAMP1-GFP (green) in WT RAW264.7 cells before and after challenge with IgG-coated microspheres. E-F) 10 kDa dextran or SRG pulsed for 16 h and chased for 1 h in WT and TMEM63A KO BMDM. G) Lysosomal pH in WT and TMEM63A KO BMDM determined using ratiometric measurements of 10 kDa Oregon Green dextran pulsed for 16 h and chased for 1 h. Each data point represents a field containing >5 cells. n = 3. H) LAMP1, cathepsin C expression and processing by WB. I) DQ-BSA signal of WT and TMEM63A KO BMDM after pulse (1 h) and chase (1 h). All data points represent a field containing 4-8 cells. n = 3. J-K) Phagocytosis in WT and TMEM63A KO BMDM, quantified in K. Each data point represents 3-5 fields containing 15-25 cells. n = 3, scale bars 10 µm.

Article Snippet: Primary antibodies against LAMP1 (DSHB, 1D4B), Cathepsin C (Santa Cruz, sc-74590) and GAPDH (Santa Cruz, 365062) were used at 1:1000 (v/v) for western blotting.

Techniques: Incubation, Expressing

Journal: bioRxiv

Article Title: Fasting reverses PLN R14del-mediated cardiomyopathy through lysosomal reactivation

doi: 10.64898/2026.03.24.713684

Figure Lengend Snippet: A. Volcano plot showing all significantly (p-value < 0.05) downregulated (n = 201) and upregulated (n = 433) genes in wild type (WT) and plna R14del zebrafish hearts with a log₂(fold change) higher than +/− 0.7. Each dot represents one gene; blue corresponds to downregulated genes and green to upregulated genes. The p-value cut off corresponds to 1.30 on the -log 10 axis. B. Fold enrichment of downregulated and upregulated GO terms identified using the DAVID online GO analysis tool, when comparing WT and plna R14del zebrafish hearts in a pseudo-bulk dataset. C. Representative images of immunofluorescent staining against DAPI, Pln and Lamp1 in one year old WT and plna R14del zebrafish. Data shown are representative for n = 8 WT and n = 8 plna R14del zebrafish hearts. Scale bar = 5 μm. White arrowheads indicate Pln and Lamp1 co-localization. D. Quantification of number of Lamp1 foci divided by cardiomyocyte (CM) area. Statistics were performed by two-tailed unpaired t-test. Data obtained from n = 8 WT and n = 8 plna R14del zebrafish hearts. E. Quantification of Pln puncta co-localized with Lamp1 foci. Data obtained from n = 8 WT and n = 8 plna R14del zebrafish hearts. F. Representative image of immunofluorescent staining against DAPI, PLN and LAMP1 in left ventricle (LV) from patients with end-stage cardiomyopathy carrying the PLN R14del variant. Dashed box high lights the zoomed in area. Data shown are representative for n = 4 individuals. Scale bars = 10 μm. White arrowheads indicate PLN and LAMP1 co-localization. G. Representative images of immunoelectron microscopy against Lamp1 (10 nm gold) in one year old WT and plna R14del zebrafish hearts. * Indicate endo-lysosomal compartments. Data shown are representative for n = 3 individuals. Scale bar = 200 nm. H. Representative images of DQ red BSA BSA ex vivo assay in one year old WT and plna R14del hearts. Dots represent functional lysosomes. Data shown are representative for n = 4 WT and n = 3 plna R14del zebrafish hearts. Scale bar = 10 μm. I. Quantification of number of DQ red BSA foci per 5000 μm 2 . Data points represent an individual measurement per image; multiple points originate from one individual. Data obtained from n = 4 WT and n = 3 plna R14del zebrafish hearts. Error bars indicate mean ± s.d. Statistics were performed by two-tailed unpaired t-test. J. Representative images of DQ red BSA ex vivo assay in WT cardiac slices treated with medium or thapsigargin (100 μM). Dots represent functional lysosomes. Data shown are representative for n = 3 per group. Scale bar = 10 μm. K. Quantification of number of DQ red BSA foci per 5000 μm 2 . Data points represent an individual measurement per image; multiple points originate from one individual. Data obtained from n = 3 zebrafish hearts per group. Error bars indicate mean ± s.d. Statistics were performed by two-tailed unpaired t-test.

Article Snippet: Sections were then blocked for 5 min with 1% BSA-c/VG and incubated for 1 hour at room temperature with a primary antibody against Lamp1 (1:10, Boster Biological Technology, DZ01398-1).

Techniques: Staining, Two Tailed Test, Variant Assay, Immuno-Electron Microscopy, Ex Vivo, Functional Assay

Journal: bioRxiv

Article Title: Phagosome Maturation in Macrophages is Enhanced by p38α MAPK Signaling

doi: 10.1101/2025.07.03.662985

Figure Lengend Snippet: A – Schematic describing the design of experiment prior to microscopy. B – Representative maximum intensity projections (MIPs) of RAW 264.7 cells at indicated time points after introduction of particles. Magenta – LAMP1, Green – 500 nm-sized non-modified polystyrene particles, Blue – Hoechst. Scale bar – 10 μm. C – Percentage of internalised particles per cell that colocalize with LAMP1 signal at indicated time points. Kruskal-Wallis test, N ≥ 3 independent experiments, 116 cells per group on average. Data represented as violin plots with colored lines indicating mean, black lines indicating median and dashed lines indicating quartiles of the data. D – Representative MIPs of RAW 264.7 cells stained with LysoTracker Red (LTR) at indicated time points after introduction of particles. Magenta – LTR, Green – 500 nm-sized non-modified polystyrene particles. Scale bar – 10 μm. E – Percentage of internalised particles per cell that colocalize with LTR signal at indicated time points, as obtained using the PCC-based method for analysis of colocalisation. Kruskal-Wallis test, N = 3 independent experiments, 55 cells per group on average. Data represented as violin plots with colored lines indicating mean, dark lines indicating median and dotted lines indicating quartiles of the data. F – Representative MIPs of RAW 264.7 cells at indicated times post infection with E. coli (MOI = 10:1). Magenta – LTR, Green – EGFP-expressing E. coli. Scale bar – 5 μm. G – Percentage of bacteria taken up by a cell that were localized in lysosomes. Kruskal-Wallis test, N = 3 independent experiments, n = 47 cells per group on average. Bars indicate the arithmetic mean of the data.

Article Snippet: Primary antibody against LAMP1 (rat anti-mouse) was procured from Developmental Studies Hybridoma Bank, Iowa (Clone number: 1D4B) and primary and secondary antibodies used for Western blot assays (rabbit anti-mouse: p38 MAPK – polyclonal, GAPDH – clone 14C10 and β tubulin – polyclonal, and goat anti-rabbit: HRP-linked anti-rabbit IgG) were purchased from Cell Signaling Technologies, USA.

Techniques: Microscopy, Modification, Staining, Infection, Expressing, Bacteria

Journal: bioRxiv

Article Title: Phagosome Maturation in Macrophages is Enhanced by p38α MAPK Signaling

doi: 10.1101/2025.07.03.662985

Figure Lengend Snippet: A – Representative MIPs of untreated cells (control) or LPS-activated RAW 264.7 cells at indicated time points post uptake of 500 nm non-modified polystyrene particles (Green) and stained with LTR (Magenta). Scale bar – 10 μm. B – Percentage colocalization of particles with LTR signal at indicated time points. Mann Whitney test; N = 3 independent experiments, 49 cells per group on average. C – Representative MIPs of primary peritoneal macrophages isolated from mice and either left untreated (control) or activated with LPS before ex vivo phagocytosis of particles. Magenta – LAMP1; Green – 500 nm-sized polystyrene particles. Scale bar – 10 μm. D – Percentage of internalized particles per cell that colocalize with LAMP1 in mouse peritoneal macrophages. Mann Whitney test; N = 3 independent experiments, 66 cells per group on average. E – Representative MIPs of RAW 264.7 cells after uptake of either control or LPS-adsorbed 500 nm-sized fluorescent particles at the indicated time points. Magenta – LysoTracker Red, Green – 500nm sized particles. Scale bars – 10 μm. F – Percentage colocalization of particles with LTR signal at indicated time points. Mann-Whitney test; N = 3 independent experiments, 94 cells per group on average. G – Representative MIPs of primary peritoneal macrophages isolated from mice after in vivo phagocytosis of particles. Magenta – LAMP1; Green – 500 nm sized particles Scale bars – 10 μm. H – Percentage of internalized particles per cell that colocalize with LAMP1 in mouse peritoneal macrophages. Mann Whitney test; N = 4 mice (control) and 3 mice (LPSads), 144 and 103 cells per group respectively. In all violin plots, dark lines represent median, and dashed lines represent quartiles of the data.

Article Snippet: Primary antibody against LAMP1 (rat anti-mouse) was procured from Developmental Studies Hybridoma Bank, Iowa (Clone number: 1D4B) and primary and secondary antibodies used for Western blot assays (rabbit anti-mouse: p38 MAPK – polyclonal, GAPDH – clone 14C10 and β tubulin – polyclonal, and goat anti-rabbit: HRP-linked anti-rabbit IgG) were purchased from Cell Signaling Technologies, USA.

Techniques: Control, Modification, Staining, MANN-WHITNEY, Isolation, Ex Vivo, In Vivo

Journal: bioRxiv

Article Title: Phagosome Maturation in Macrophages is Enhanced by p38α MAPK Signaling

doi: 10.1101/2025.07.03.662985

Figure Lengend Snippet: A – Representative MIPs of RAW 264.7 cells 2 hours after uptake of either control particles or particles conjugated with IgG, albumin or folic acid. Magenta – LAMP1, Green – particles. Scale bars – 5 μm. B – Percentage of phagocytosed particles per cell that colocalize with LAMP1 signal in RAW264.7 cells. Kruskal-Wallis test; N = 3 independent experiments, 130 cells per group on average. C – Representative MIPs of primary peritoneal macrophages isolated from mice 2h after ex vivo internalization of either control particles or particles conjugated with either mouse IgG, albumin or folic acid. Magenta – LAMP1; Green – 500 nm-sized polystyrene particles. Scale bars – 10 μm. D – Percentage of phagocytosed particles per cell that colocalize with LAMP1 signal in mouse peritoneal macrophages. Kruskal-Wallis test; N = 3 independent experiments, 58 cells per group on average. In all violin plots, dark lines represent median and dashed lines represent quartiles of the data.

Article Snippet: Primary antibody against LAMP1 (rat anti-mouse) was procured from Developmental Studies Hybridoma Bank, Iowa (Clone number: 1D4B) and primary and secondary antibodies used for Western blot assays (rabbit anti-mouse: p38 MAPK – polyclonal, GAPDH – clone 14C10 and β tubulin – polyclonal, and goat anti-rabbit: HRP-linked anti-rabbit IgG) were purchased from Cell Signaling Technologies, USA.

Techniques: Control, Isolation, Ex Vivo

Journal: bioRxiv

Article Title: Phagosome Maturation in Macrophages is Enhanced by p38α MAPK Signaling

doi: 10.1101/2025.07.03.662985

Figure Lengend Snippet: A – Representative MIPs of RAW 264.7 cells 2 hours post uptake of particles. Cells were treated with p38MAPK inhibitor SB203580 (10 μM) or vehicle (DMSO) for 1h followed by 100 ng/mL LPS for 18h before uptake, or left untreated (control). Magenta – LAMP1, Green – 500 nm polystyrene particles. Scale bars – 10 μm. B – Percentage of particles taken up per cell that colocalize with LAMP1 signal 2 hours post uptake in RAW 264.7 cells after treatments described in A. Kruskal-Wallis (with Dunn’s post-hoc) test; N = 3 independent experiments, n = 219 cells per group on average. C – Representative MIPs of RAW 264.7 cells which were treated with either 10 μM SB203580 or vehicle (DMSO) or received no treatment before internalization of either non-modified (control) or LPS ads particles (all other groups). Treatments occurred 2 hours-post addition of particles. Magenta – LAMP1, Green – 500 nm polystyrene particles (either non-modified – control or LPS adsorbed). Scale bars – 10 μm. D – Percentage of particles taken up per cell that localize to lysosomes, 2h post uptake after treatments described in C. Kruskal-Wallis (with Dunn’s post-hoc) test; N ≥ 3 independent experiments, n = 226 cells on average per group. Data are plotted as violin plots with black lines indicating median, dashed lines indicating quartiles, and colored lines indicating mean of the data.

Article Snippet: Primary antibody against LAMP1 (rat anti-mouse) was procured from Developmental Studies Hybridoma Bank, Iowa (Clone number: 1D4B) and primary and secondary antibodies used for Western blot assays (rabbit anti-mouse: p38 MAPK – polyclonal, GAPDH – clone 14C10 and β tubulin – polyclonal, and goat anti-rabbit: HRP-linked anti-rabbit IgG) were purchased from Cell Signaling Technologies, USA.

Techniques: Control, Modification

Journal: Journal of Orthopaedic Translation

Article Title: Activation of LAMP1-mediated lipophagy by sulforaphane inhibits cellular senescence and intervertebral disc degeneration

doi: 10.1016/j.jot.2025.05.010

Figure Lengend Snippet: LAMP1 suppressed lipid accumulation and senescence in NP cells. ( A and B ) Immunohistochemical detection of LAMP1 expression in mildly and severely degenerated NP tissues, scale bar = 100 μM. ( C and D ) LAMP1 expression in normal and degenerated rat NP tissues, scale bar = 500 μM. ( E ) Triglyceride content in rat NP cells after PA treatment and LAMP1 overexpression. ( F ) Cholesterol ester content in rat NP cells after PA treatment and LAMP1 overexpression. ( G and H ) Expression of LAMP1 and PLIN2 in rat NP cells after PA treatment and LAMP1 overexpression, detected by Western blot. ( I ) LDs in rat NP cells after PA treatment and LAMP1 overexpression, observed with Biodipy staining, scale bar = 50 μM. ( J ) Senescence of rat NP cells after PA treatment and LAMP1 overexpression, detected by β-galactosidase staining, scale bar = 100 μM. ( K and L ) Expression of senescence markers (p53, p21, and p16) in rat NP cells after PA treatment and LAMP1 overexpression, detected by Western blot. Data are represented as mean ± SD, ∗ p < 0.05.

Article Snippet: Primary antibodies against LAMP1 (Santa Cruz, sc-20011, USA) and LC3 (Proteintech, 14600-1-AP, USA) were added to the supernatants, and the mixture was rotated overnight at 4 °C.

Techniques: Immunohistochemical staining, Expressing, Over Expression, Western Blot, Staining

Journal: Journal of Orthopaedic Translation

Article Title: Activation of LAMP1-mediated lipophagy by sulforaphane inhibits cellular senescence and intervertebral disc degeneration

doi: 10.1016/j.jot.2025.05.010

Figure Lengend Snippet: SFN promoted LAMP1 expression and lipophagy. ( A ) Chemical structure of SFN. ( B and C ) Expression of LAMP1 and autophagy markers (P62 and LC3) in rat NP cells after PA and SFN treatment, detected by WB. ( D and E ) LAMP1 expression in rat NP cells after PA and SFN treatment, observed by immunofluorescence, scale bar = 50 μM. ( F and G ) LC3 expression in rat NP cells after PA and SFN treatment, observed by fluorescence microscopy, scale bar = 50 μM. ( H ) Interaction between LAMP1 and LC3 in rat NP cells after PA and SFN treatment, detected by Co-IP. ( I ) LDs in rat NP cells after PA and SFN treatment, observed by TEM, the red arrow indicates the autolysosomes, scale bar = 5 μM. ( J and K ) Co-localization of LAMP1 and LDs in rat NP cells after PA and SFN treatment, observed by fluorescence microscopy, scale bar = 30 μM. ( L and M ) Co-localization of LC3 and LDs in rat NP cells after PA and SFN treatment, observed by fluorescence microscopy, scale bar = 30 μM. Data are represented as mean ± SD, ∗ p < 0.05.

Article Snippet: Primary antibodies against LAMP1 (Santa Cruz, sc-20011, USA) and LC3 (Proteintech, 14600-1-AP, USA) were added to the supernatants, and the mixture was rotated overnight at 4 °C.

Techniques: Expressing, Immunofluorescence, Fluorescence, Microscopy, Co-Immunoprecipitation Assay

Journal: Journal of Orthopaedic Translation

Article Title: Activation of LAMP1-mediated lipophagy by sulforaphane inhibits cellular senescence and intervertebral disc degeneration

doi: 10.1016/j.jot.2025.05.010

Figure Lengend Snippet: SFN delayed senescence in NP cells through LAMP1-mediated lipophagy. ( A ) Triglyceride content in rat NP cells after PA, SFN treatment, and siRNA-LAMP1 transfection. ( B ) Cholesterol ester content in rat NP cells after PA, SFN treatment, and siRNA-LAMP1 transfection. ( C and D ) Expression of LAMP1 and PLIN2 in rat NP cells after PA, SFN treatment, and siRNA-LAMP1 transfection, detected by Western blot. ( E ) LDs in rat NP cells after PA, SFN treatment, and siRNA-LAMP1 transfection, observed with Biodipy staining, scale bar = 50 μM. ( F ) Senescence and proliferation of rat NP cells after PA, SFN treatment, and siRNA-LAMP1 transfection, scale bar = 100 μM (white light images), scale bar = 500 μM (IF images). ( G and H ) Cell cycle distribution of rat NP cells after transfection with LAMP1 siRNA, analyzed by flow cytometry, ∗p < 0.05 compared with CTR group; #p < 0.05 compared with PA group; &p < 0.05 compared with PA + SFN group. ( I-L ) Expression of senescence markers in rat NP cells after PA, SFN treatment, and siRNA-LAMP1 transfection, detected by Western blot. Data are represented as mean ± SD, ∗ p < 0.05.

Article Snippet: Primary antibodies against LAMP1 (Santa Cruz, sc-20011, USA) and LC3 (Proteintech, 14600-1-AP, USA) were added to the supernatants, and the mixture was rotated overnight at 4 °C.

Techniques: Transfection, Expressing, Western Blot, Staining, Flow Cytometry

Journal: Journal of Orthopaedic Translation

Article Title: Activation of LAMP1-mediated lipophagy by sulforaphane inhibits cellular senescence and intervertebral disc degeneration

doi: 10.1016/j.jot.2025.05.010

Figure Lengend Snippet: SFN attenuated lipid accumulation and senescence in a rat IDD model. ( A ) Schematic diagram of the experimental protocol in SD rats. ( B ) MRI and X-ray images of rat caudal vertebrae. ( C ) Assessment of rat IDD using the Pfirrmann grading system. ( D ) Evaluation of changes in intervertebral disc height (DHI) in rats. ( E and F ) Evaluation and scoring of rat IDD using HE staining and Safranin O-Fast Green staining, scale bar = 500 μM. ( G-J ) Immunohistochemical detection of LAMP1, PLIN2, and p21 expression in rat NP tissue, scale bar = 500 μM. ( K ) Immunofluorescence detection of LAMP1 and PLIN2 expression in rat NP tissue, scale bar = 500 μM. Data are represented as mean ± SD, ∗ p < 0.05.

Article Snippet: Primary antibodies against LAMP1 (Santa Cruz, sc-20011, USA) and LC3 (Proteintech, 14600-1-AP, USA) were added to the supernatants, and the mixture was rotated overnight at 4 °C.

Techniques: Staining, Immunohistochemical staining, Expressing, Immunofluorescence