lamp1 antibody  (Developmental Studies Hybridoma Bank)

Journal: The Journal of Cell Biology

Article Title: Rsp5/NEDD4 and ESCRT regulate TDP-43 toxicity and turnover via an endolysosomal clearance mechanism

doi: 10.1083/jcb.202212064

Figure Lengend Snippet: Mild TDP-43 overexpression causes accumulation of giant Rab7-positive vesicles. (A) HEK293A WT, HEK-TDP-43-GFP, and HEK-TDP-35-GFP cells were grown in normal conditions before being fixed and immunostained for Rab7. N = 3; scale bar, 10 µm. (B) Quantification of the frequency of giant endosomes in HEK293A WT, HEK-TDP-43-GFP, and HEK-TDP-35-GFP cells. **, P < 0.01; ***, P < 0.001 by one-way analysis of variance with Tukey’s post hoc test; N = 3. (C) Quantification of the average size of giant endosomes in HEK293A WT, HEK-TDP-43-GFP, and HEK-TDP-35-GFP cells. ***, P < 0.001 by one-way analysis of variance with Tukey’s post hoc test; N = 3. (D) Distribution of giant Rab7+ve vesicle diameters from WT, HEK-TDP-43-GFP, and HEK-TDP-35-GFP cell lines. N = 3. (E) HEK-TDP-43-GFP (pseudocolored white) cells were grown in normal conditions before being fixed and immunostained for Rab5, LC3B, or LAMP1 (pseudocolored green) and then successively immunostained for Rab7 (pseudocolored red). N = 3; scale bar, 10 µm. (F) HEK-TDP-35-GFP (pseudocolored white) cells were grown in normal conditions before being fixed and immunostained for Rab5, LC3B, or LAMP1 (pseudocolored green) and then successively immunostained for Rab7 (pseudocolored red). N = 3; scale bar, 10 µm in main panels; 2 µm in insets. Error bars in all graphical panels represent standard error of the mean.

Article Snippet: Primary antibodies used for the immunofluorescence were Rab7 (ab137029; Abcam, dilution: 1:200), CD63 (H5C6; Developmental Studies Hybridoma Bank, dilution 1:100), Rab5 (ab109534; Abcam, dilution: 1:200), LC3B (3868S; Cell Signaling, dilution 1:500), LAMP1 (21997-1-AP; Proteintech, dilution 1:200), and TDP-43 (10782-2-AP; Proteintech; dilution 1:200).

Techniques: Over Expression

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