anti rab7 antibody  (Proteintech)

Journal: International Journal of Molecular Medicine

Article Title: Macrophage SH3 domain-containing GRB2-like protein B1 is required for Rab7-mediated mitophagy in inflammation during sea water drowning acute lung injury

doi: 10.3892/ijmm.2026.5809

Figure Lengend Snippet: SH3GLB1 interacts with the lysosomal protein Rab7 to contribute to macrophage inflammation. (A) Representative top five proteins potentially binding to SH3GLB1 in mitochondria identified by mass spectrometry analysis. (B) Representative western blots of Rab7 protein levels in BMDMs treated with LPS or sea water. (C) Co-IP assay showing the interaction of SH3GLB1 with Rab7. (D) Representative images of immunofluorescence staining for SH3GLB1 (green) and Rab7 (red) in BMDMs. Scale bar, 20 µ m. (E) Representative molecular docking analysis of SH3GLB1 and Rab7. (F) Western blots of recombinant Rab7 binding to recombinant SH3GLB1. Representative ELISA results showing the levels of inflammatory factors, including IL-1β, IL-6 and TNFα, in the supernatants of cells treated with (G and H) LPS (n=3) or (I and J) sea water (n=3). The data are presented as the means ± SEMs; P<0.05 was considered to indicate statistical significance. SH3GLB1, SH3 domain-containing GRB2-like protein B1; BMDMs, bone marrow-derived macrophages; LPS, lipopolysaccharide; Co-IP, coimmunoprecipitation.

Article Snippet: Primary antibodies: GAPDH (1:1,000; cat. no. ab8245; Abcam), HSP90 (1:1,000; cat. no. 13171-1-AP, Proteintech Group, Inc.), SH3GLB1 (1:1,000; cat. no. sc-374146, Santa Cruz Biotechnology, Inc.), LC3 (1:1,000; cat. no. 12741T; CST Biological Reagents Co., Ltd.), P62 (1:1,000; cat. no. ab109012, Abcam), Rab7 (1:1,000; cat. no. A12308, ABclonal) and Parkin (1:1,000; cat. no. ab77924, Abcam).

Techniques: Binding Assay, Mass Spectrometry, Western Blot, Co-Immunoprecipitation Assay, Immunofluorescence, Staining, Recombinant, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: International Journal of Molecular Medicine

Article Title: Macrophage SH3 domain-containing GRB2-like protein B1 is required for Rab7-mediated mitophagy in inflammation during sea water drowning acute lung injury

doi: 10.3892/ijmm.2026.5809

Figure Lengend Snippet: SH3GLB1 facilitated adverse Rab7-mediated mitophagy. (A) Representative images of immunofluorescence staining for SH3GLB1 (green) and mitochondria (red) or lysosomes (red) in BMDMs. Scale bar, 20 µ m. Representative (B and F) western blot and (C-E and G-I) quantification of mitophagy-related protein levels in BMDMs treated with CCCP. (J) mKeima expressed in BMDMs was imaged by confocal microscopy (mKeima-561 nm: red; mKeima-488 nm: green; scale bar, 20 µ m); (K) the 561/488 nm ratio of mKeima was statistically quantified (n=3). (L) Representative flow cytometry analysis of mKeima in BMDMs. The data are presented as the means ± SEMs; P<0.05 was considered to indicate statistical significance. SH3GLB1, SH3 domain-containing GRB2-like protein B1; BMDMs, bone marrow-derived macrophages; CCCP, carbonyl cyanide-chlorophenylhydrazone.

Article Snippet: Primary antibodies: GAPDH (1:1,000; cat. no. ab8245; Abcam), HSP90 (1:1,000; cat. no. 13171-1-AP, Proteintech Group, Inc.), SH3GLB1 (1:1,000; cat. no. sc-374146, Santa Cruz Biotechnology, Inc.), LC3 (1:1,000; cat. no. 12741T; CST Biological Reagents Co., Ltd.), P62 (1:1,000; cat. no. ab109012, Abcam), Rab7 (1:1,000; cat. no. A12308, ABclonal) and Parkin (1:1,000; cat. no. ab77924, Abcam).

Techniques: Immunofluorescence, Staining, Western Blot, Confocal Microscopy, Flow Cytometry, Derivative Assay

Journal: medRxiv

Article Title: Biallelic WDR91 variants cause a neurodevelopmental disorder through impaired endosomal maturation and autophagy dysregulation

doi: 10.64898/2026.04.03.26349989

Figure Lengend Snippet: A. Time-lapse recording of dynamic changes of the early endosomal marker 2×FYVE-mCherry and the late endosomal protein GFP-Rab7 on endosomes in transduced HEK193 cells (left) and duration of the overlap of 2×FYVE and Rab7 on endosomes (right; one-way ANOVA with Kruskal-Wallis multiple comparison test, ** p<0.01, **** p<10 - ; at least 10 endosomes per cell line were analyzed). B. Colocalization of endogenous EEA1 with Rab7 in WDR91 WT and WDR91 Y15N cells: representative confocal images (left) and assessment of Pearson’s coefficient (right; Mann-Whitney test, *** p<10 - ; at least 30 independent cells per cell line were analyzed). C. EEA1 and Rab7 protein expression in transduced HEK293 cells, representative blot of three independent experiments. Individual fluorescence channels were analyzed separately due to distinct background signal. D. Colocalization of WDR91 with endogenous EEA1 or Rab7 in WDR91 WT and WDR91 Y15N cells: representative confocal images (left) and assessment of Pearson’s coefficient (right; 2-way ANOVA with Sidak multiple comparison test, *** p<10 -3 , **** p<10 -4 ; at least 30 independent cells per cell line were analyzed).

Article Snippet: Cells were then saturated using TBS-Tween 0.01% supplemented with 3% goat serum, and stained with a primary anti-human WDR91 Ab (Abcam) and anti-human EEA1 or anti-human Rab7 Abs (Cell Signaling Technologies) and a secondary goat anti-mouse/rabbit IgG mAb coupled to AF488 or AF687 (Thermo Fisher Scientific).

Techniques: Marker, Comparison, MANN-WHITNEY, Expressing, Fluorescence

Journal: Redox Biology

Article Title: YY1 nitration participates in DbCM cardiomyocyte lipotoxicity by inhibiting ANXA3 -induced microlipophagy

doi: 10.1016/j.redox.2026.104085

Figure Lengend Snippet: ANXA3 participated in T2DM-induced DbCM by regulating microlipophagy. (A-B) Representative images of TEM were used to observe the autophagy and microlipophagy in the hearts of mice. The red arrow represents autophagosomes, and the yellow arrow represents lipid droplets in contact with lysosomes. Bars: 1 μm (C) Relative Rab7a mRNA expression levels in heart tissues of mice analyzed by RT-qPCR, n = 4. (D-E) Relative Rab7a protein expression levels in the heart of mice analyzed by Western blot, n = 4. (F) Relative RAB7A mRNA expression levels in AC16 cells analyzed by RT-qPCR, n = 3-6. (G-H) Relative Rab7a protein expression levels in AC16 cells analyzed by Western blot, n = 3-5. The data were presented as the mean ± SD. ∗ P < 0.05 versus the db/db + OE-GFP group or the si-NC group; # P < 0.05 versus the OE-NC group.

Article Snippet: Blocked with 5% (w/v) non-fat-dried milkat room temperature for 1 h. Then the membranes were incubated with the anti-ANXA3 antibody (Proteintech, 11804-1-AP; 1:1000 [v/v]), the anti-YY1 antibody (Proteintech, 22156-1-AP; 1:1000 [v/v]) the anti-PLIN2 antibody (Proteintech, 15294-1-AP; 1:1000 [v/v]), the anti-SQSTM1/p62 antibody (Cell Signaling Technology, 23214; 1:1000 [v/v]), the anti-LC3 antibody (Cell Signaling Technology, 12741; 1:1000 [v/v]), the anti-Rab7 antibody (Proteintech, 55469-1-AP; 1:1000 [v/v])or the rabbit anti-α-Tubulin antibody (ABclonal, AC031; 1:1000 [v/v])overnight at 4 °C.

Techniques: Expressing, Quantitative RT-PCR, Western Blot