lamp2 rat mab  (Developmental Studies Hybridoma Bank)

Journal: Acta Neuropathologica Communications

Article Title: Pathobiology of the autophagy-lysosomal pathway in the Huntington’s disease brain

doi: 10.1186/s40478-025-02131-8

Figure Lengend Snippet: Correlation between marker expression levels and HD progression stages across different brain regions, grouped by marker category. The heatmap illustrates Spearman’s rank correlation coefficients (ρ) between the relative mRNA expression levels from qPCR of selected molecular markers and Huntington’s disease (HD) severity. Analyses were performed on human post-mortem brain tissue from control (Ctl) and HD patients (grades HD2, HD3, HD4), with severity numerically encoded for correlation (Ctl = 0, HD2 = 1, HD3 = 2, HD4 = 3). Correlations are shown for STR, CTX, and CBM). Markers are grouped into functional categories, displayed from top to bottom: Induction Markers (Autophagy induction): ATG7, LC3, p62. Lysosomal Markers (Lysosomal biogenesis, hydrolases and structural elements): TFEB, TFE3, CTSB, CTSD, HEXA, LAMP1, LAMP2. Cellular Markers (cellular processes): ENO2, TUBB3, GFAP, AIF1, MBP. The color of each cell represents the Spearman ρ value, with a continuous gradient from blue (strong negative correlation, ρ = −1) through white (no correlation, ρ = 0) to red (strong positive correlation, ρ = + 1). Nominal p-values for the correlations are indicated by asterisks: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Antibodies for immunohistochemistry (IHC), western blotting (WB): The following primary antibodies were used in this study. (1) from Cell Signaling Technology: tHTT rabbit mAb (clone D7F7, #5656, targeting residues surrounding Pro1220 of human HTT and detecting total HTTs), p70S6K pAb (#9202), p-p70S6K (T389) pAb (#9205), ULK1 pAb (#4773), p-ULK1 (S757) pAb (#6888, #14202; detecting S757 or S758 of mouse or human ULK1, respectively), ATG5 rabbit mAb (#12994), ATG7 pAb (#2631), ATG13 rabbit mAb (#13273), p-ATG13 (S355) rabbit mAb (#26839), VPS34 rabbit mAb (#81453), TRAF6 rabbit mAb (#8028), Calnexin rabbit mAb (#2679). (2) from Millipore-Sigma: ntHTT mAb (N-Terminus-specific, mEM48, #MAB5374, preferentially recognizing aggregated HTT) , ATG5 pAb (#ABC14), K48- or K63-specific ubiquitin mAb (#05–1307, #05–1308, respectively), βIII-tubulin mAb (#SAB4700544), β–actin mAb (#A1978). (3) from other vendors: BECN1 mAb (BD Biosciences, #612113); LC3 pAb (Novus Biologics, #NB100-2220), ATG9 (Novus Biologics, #B-110–56893); p62 mAb (BD Biosciences, #610832) or C-term-specific p62 Guinea Pig pAb (Progen Biotechnik, #C-1620); total ubiquitin pAb (Dako Agilent, #Z0458), LAMP1 or LAMP2 rat mAb (Developmental Studies Hybridoma Bank, University of Iowa, #H4A3 or #H4B4, respectively); CTSD sheep pAb (D-2–3, in-house made) ( ); CTSD pAb (Scripps Laboratories, #RC245), CTSD mAb (CD1.1, in-house made) ( ); CTSB pAb (Cortex Biochemicals, #CR6009RP), CTSB goat pAb (Neuromics, #GT15047).

Techniques: Marker, Expressing, Control, Functional Assay

Journal: Frontiers in Immunology

Article Title: SFB flagellin mediates cell adhesion, endocytosis and immune regulation in germ-free mice

doi: 10.3389/fimmu.2025.1624092

Figure Lengend Snippet: Co-localization analysis of SFB rFliC3 with endosomal marker LAMP2. Ileal tissues were fixed in 4% paraformaldehyde and sequentially immune-stained with rabbit anti-SFB FliC3 primary antibody, anti-LAMP2 antibody, and FITC-conjugated goat anti-rabbit IgG secondary antibody. Nuclei were counterstained with DAPI. Fluorescence imaging was performed using an upright microscope (Zeiss Axio Imager Z2). (A) DAPI staining results of ileal tissue; (B) Anti-SFB FliC3 antibody staining results of ileal tissue; (C) Anti-LAMP2 antibody staining results of ileal tissue; (D) Merged images of (A–C) demonstrating co-localization signals (yellow) of FliC3 and LAMP2.

Article Snippet: Given that SFB antigens were internalizes into cells through endocytosis to exert immune regulatory effects, and the lysosome marker LAMP2 associated with cell adhesion is involved in this process , we investigated the endocytic capacity of SFB fliC3 -expressing Lactococcus by using dual immunofluorescence staining with anti-SFB FliC3 polyclonal antibody and anti- Lamp2 monoclonal antibody (supplied by DSHB, ABL-93).

Techniques: Marker, Staining, Fluorescence, Imaging, Microscopy

Journal: Frontiers in Immunology

Article Title: SFB flagellin mediates cell adhesion, endocytosis and immune regulation in germ-free mice

doi: 10.3389/fimmu.2025.1624092

Figure Lengend Snippet: Subcellular localization of mFliC3 in MODE-K murine intestinal epithelial cells. MODE-K cells were incubated with mfliC3 -expressing E. coli (BW25113 ΔfliC : tac-mfliC3 ) for 12 hours. Following PBS washing, triple immunofluorescence staining was performed using DAPI nuclear counterstain (blue), anti-mFliC3 antibody (green), and anti-LAMP2 endosomal marker (red). (A) DAPI staining results of cells; (B) Anti-SFB FliC3 antibody staining results of cells; (C) Anti-LAMP2 antibody staining results of cells; (D) Merged images of (A–C) demonstrating co-localization signals (yellow) of FliC3 and LAMP2. (D) shows merged fluorescence channels. Specimens were mounted in anti-fade medium and imaged by confocal microscopy (scale bar: 10 μm).

Article Snippet: Given that SFB antigens were internalizes into cells through endocytosis to exert immune regulatory effects, and the lysosome marker LAMP2 associated with cell adhesion is involved in this process , we investigated the endocytic capacity of SFB fliC3 -expressing Lactococcus by using dual immunofluorescence staining with anti-SFB FliC3 polyclonal antibody and anti- Lamp2 monoclonal antibody (supplied by DSHB, ABL-93).

Techniques: Incubation, Expressing, Immunofluorescence, Staining, Marker, Fluorescence, Confocal Microscopy

Journal: Biology

Article Title: FAM98 Family Proteins Play Distinct Roles in Osteoclastogenesis and Bone Resorption

doi: 10.3390/biology14010045

Figure Lengend Snippet: The key reagents used in this study.

Article Snippet: , Rat monoclonal anti-Lamp-2 , Developmental Studies Hybridoma Bank, Iowa City, IA, USA , GL2A7 , IF 1:200.

Techniques: Red Blood Cell Lysis, Plasmid Preparation, Transduction, Transfection, Staining, Lysis, Protease Inhibitor, Purification, Gene Expression, Quantitative RT-PCR