mouse anti prkn monoclonal antibody  (Santa Cruz Biotechnology)

Journal: Cell & Bioscience

Article Title: PINK1 functions in mitophagy and mitochondrial homeostasis during mice oocyte maturation

doi: 10.1186/s13578-025-01460-4

Figure Lengend Snippet: RAB8A is required for PINK1-mediated mitophagy in murine oocytes. ( A ) The effects of Pink1 -OE on PRKN and RAB8A protein levels, as detected by Western blot. Proteins from 300 oocytes were loaded for every sample. ( B ) Relative intensity results for the protein expression of PRKN in control and Pink1 -OE oocytes. ( C ) Relative intensity results for the protein expression of RAB8A in control and Pink1 -OE oocytes. ( D ) The effects of 10 μm CCCP on PINK1, PRKN, and RAB8A protein levels, as detected by Western blot. ( E ) Relative intensity results for the protein expression of PINK1 in control and 10 μm CCCP treated oocytes. ( F ) Relative intensity results for the protein expression of PRKN in control and 10 μm CCCP treated oocytes. ( G ) Relative intensity results for the protein expression of RAB8A in control and 10 μm CCCP treated oocytes. Proteins from 300 oocytes were loaded for every sample

Article Snippet: Rabbit monoclonal anti-γH2AX antibody (Cat#: ab81299), rabbit monoclonal anti-RAB8A antibody (Cat#: ab188574), mouse monoclonal anti-Myc antibody (Cat#: ab18185) were purchased from Abcam (Cambridge, UK); rabbit polyclonal anti-PINK1 antibody (Cat#: PA5-85243) was obtained from Thermo Fisher Scientific (Thermo Fisher Scientific, USA); mouse monoclonal- FITC-conjugated anti-α-Tubulin antibody (Cat#: F2168) was bought from Sigma (St. Louis, MO, USA); rabbit polyclonal antibodies anti-LC3 antibody (Cat#: 14600-1-AP), mouse-Monoclonal antibody (Cat#: 60004-1-Ig), rabbit polyclonal antibody anti-α-Tubulin antibody (Cat#: 11224-1-AP) were purchased from Proteintech (Wuhan, China); mouse monoclonal antibody anti-PRKN antibody (Cat#: sc-32282) was obtained from Santa Cruz Biotechnologies (Santa Cruz, CA).

Techniques: Western Blot, Expressing, Control

Journal: Cell & Bioscience

Article Title: PINK1 functions in mitophagy and mitochondrial homeostasis during mice oocyte maturation

doi: 10.1186/s13578-025-01460-4

Figure Lengend Snippet: RAB8A activity is essential for murine oocyte quality in advanced maternal age. (A) The effects of advanced maternal age on RAB8A protein level. (B) The effects of advanced maternal age on PRKN protein level. (C) Western blotting with anti-Myc showed that Rab8a - T22N-Myc and Rab8a - Q67L-Myc were efficiently overexpressed. (D) Ultrastructure observation of Young, Old, Old + Rab8a - WT, Old + Rab8a - T22N, Old + Rab8a - Q67L oocytes. Mitophagosomes (red arrows), healthy mitochondria (white arrows), damaged mitochondria (blue arrows). (E) Number of mitophagosomes per cell section in Young, Old, Old + Rab8a – WT, Old + Rab8a – T22N, Old + Rab8a – Q67L oocytes (1.07 ± 0.21; 1.13 ± 0.17; 1.23 ± 0.34; 1.10 ± 0.16; 2.13 ± 0.21, respectively ). Bar graphs are mean ± SD of results from 3 oocytes section. Bars: 500 nm. (The bars labeled with completely different letters indicate significant difference at P < 0.05). (F) A proposed model showing the potential pathway that RAB8A activity is involved in PINK1-PRKN mediated mitophagy in oocytes

Article Snippet: Rabbit monoclonal anti-γH2AX antibody (Cat#: ab81299), rabbit monoclonal anti-RAB8A antibody (Cat#: ab188574), mouse monoclonal anti-Myc antibody (Cat#: ab18185) were purchased from Abcam (Cambridge, UK); rabbit polyclonal anti-PINK1 antibody (Cat#: PA5-85243) was obtained from Thermo Fisher Scientific (Thermo Fisher Scientific, USA); mouse monoclonal- FITC-conjugated anti-α-Tubulin antibody (Cat#: F2168) was bought from Sigma (St. Louis, MO, USA); rabbit polyclonal antibodies anti-LC3 antibody (Cat#: 14600-1-AP), mouse-Monoclonal antibody (Cat#: 60004-1-Ig), rabbit polyclonal antibody anti-α-Tubulin antibody (Cat#: 11224-1-AP) were purchased from Proteintech (Wuhan, China); mouse monoclonal antibody anti-PRKN antibody (Cat#: sc-32282) was obtained from Santa Cruz Biotechnologies (Santa Cruz, CA).

Techniques: Activity Assay, Western Blot, Labeling

Journal: Molecular cell

Article Title: An Epstein-Barr virus protein interaction map reveals NLRP3 inflammasome evasion via MAVS UFMylation

doi: 10.1016/j.molcel.2023.05.018

Figure Lengend Snippet:

Article Snippet: Anti-PARK2 mouse monoclonal antibody (Prk8) , Cell Signaling Technology , cat# 4211; RRID: AB_2159920.

Techniques: Recombinant, Magnetic Beads, Transfection, Sequencing, Modification, Mass Spectrometry, Produced, Protease Inhibitor, Selection, Purification, Gel Extraction, SYBR Green Assay, Reporter Assay, Protein Quantitation, CRISPR, Amplification, Molecular Cloning, Plasmid Preparation, Software

Journal: Oncotarget

Article Title: Multiple-level validation identifies PARK2 in the development of lung cancer and chronic obstructive pulmonary disease

doi: 10.18632/oncotarget.9954

Figure Lengend Snippet: A-C. Cytokines, IL-6, IL-1β or TNF-α protein or mRNA levels in serum from C57BL/6 mice (A), mouse primary bronchus epithelial cells (B), MEFs (C) of PARK2 WT or KO were analyzed by ELISA or qRT-PCR. D. After infection with the indicated plasmids, cytokines in the cultured MEFs supernatants were determined by ELISA. Results are shown as means (±SEM), and at least three experiments were performed for all experiments. ***, p < 0.001 by one-way ANOVA. E. PARK2 WT and KO mice ( n=6 ) at 10 months of age were analyzed for tumor event by immunohistochemistry. H&E staining; x100 and x400.

Article Snippet: Mouse monoclonal antibody recognizing PARK2 (sc-32282) and Lamin B1 (sc-20682) were purchased from Santa Cruz Biotechnology.

Techniques: Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Infection, Cell Culture, Immunohistochemistry, Staining

Journal: Oncotarget

Article Title: Multiple-level validation identifies PARK2 in the development of lung cancer and chronic obstructive pulmonary disease

doi: 10.18632/oncotarget.9954

Figure Lengend Snippet: A. PARK2 deficiency results in centrosome amplification in MEFs at passage 5. ( left ) Immunofluorescence staining shows impaired mitoses in MEFs depleted of PARK2 . Red, γ-tubulin; White arrows, centrosome in mitosis. The scale bar is 20 μm. ( right ) Quantification of 2 < centrosomes cells. ***, p < 0.001 by one-way ANOVA. n =10. B. After infection with the indicated plasmids, MEFs lysates were fixed for immunofluorescence assay. Quantification of 2 < centrosomes cells using γ-tubulin. Results are shown as means (±SEM), and at least three experiments were performed for all experiments. *, p < 0.05 and **, p < 0.01 by one-way ANOVA. C. Immunofluorescence staining shows impaired mitoses in IMR 90 lung fibroblast cells depleted of PARK2 . White arrows, centrosome in mitosis. Yellow arrows, abnormal chromosome. The scale bar is 20 μm. D. Cell were harvested at indicated times and analyzed by immunobloting. E. Impaired mitoses including multipolar spindles, misalignment and abnormal microtubule in PARK2 knockout (KO) MEF cells. Yellow arrows, abnormal chromosome. The scale bar is 20 μm. Results are shown as means (±SEM), and at least three experiments were performed for all experiments. **, p < 0.01 by one-way ANOVA.

Article Snippet: Mouse monoclonal antibody recognizing PARK2 (sc-32282) and Lamin B1 (sc-20682) were purchased from Santa Cruz Biotechnology.

Techniques: Amplification, Immunofluorescence, Staining, Infection, Western Blot, Knock-Out

Journal: Oncotarget

Article Title: Multiple-level validation identifies PARK2 in the development of lung cancer and chronic obstructive pulmonary disease

doi: 10.18632/oncotarget.9954

Figure Lengend Snippet: A. Primary MEFs were applied on 3-D cultures system for 2 weeks and counted the sphere of the cells. ( top ) Representative images indicate the sphere cells. ( bottom ) Quantification of the sphere number. ****, p < 0.0001 by one-way ANOVA. n =6. ( B. and C. ) PARK2 WT and KO MEFs were analyzed for foci formation at passage 5 and 21 (B), and colony formation at passage 21 C. The scale bar is 20 μm. Results are shown as means (±SEM), and at least three experiments were performed for all experiments. NAC, ROS scavenger. ***, p < 0.001 by one-way ANOVA. D. The growth of lung cancer cell lines (H1437, H522, H1650, A549, H460, H1299 and H196) was inhibited through infection with PARK2 . Cells were transfected with the indicated plasmids, and then cell lysates were blotted with the indicated antibodies. E. The percentage of lung cancer cell growth compared to controls after transfected PARK2 . Results shown as means (± SEM), and at least three experiments were performed for all experiments. **, p < 0.01 and ***, p < 0.001 by one-way ANOVA. F. PARK2 and β–actin expression was measured by immunoblot.

Article Snippet: Mouse monoclonal antibody recognizing PARK2 (sc-32282) and Lamin B1 (sc-20682) were purchased from Santa Cruz Biotechnology.

Techniques: Infection, Transfection, Expressing, Western Blot

Journal: Oncotarget

Article Title: Multiple-level validation identifies PARK2 in the development of lung cancer and chronic obstructive pulmonary disease

doi: 10.18632/oncotarget.9954

Figure Lengend Snippet: Significant results of Testing 114 Informative PARK2 SNPs

Article Snippet: Mouse monoclonal antibody recognizing PARK2 (sc-32282) and Lamin B1 (sc-20682) were purchased from Santa Cruz Biotechnology.

Techniques:

Journal: Molecular and Cellular Biology

Article Title: HSPA1A-Independent Suppression of PARK2 C289G Protein Aggregation by Human Small Heat Shock Proteins

doi: 10.1128/MCB.00698-14

Figure Lengend Snippet: Suppression of PARK2 C289G aggregation by HSPBs. (A) HEK293 cells were transfected for 24 h with Flag-tagged PARK2 WT or PARK2 C289G. Subcellular distribution of PARK2 (green) was investigated by immunofluorescence. The diagram shows the percentage of PARK2 aggregates. (B) Cells, transfected as in panel A, were fractionated in TX-100-soluble and -insoluble proteins and analyzed by Western blotting. (C) HEK293 cells were transfected for 24 h with Flag-tagged PARK2 C289G alone or together with each of the different HSPB proteins. Cells were fractionated in TX-100-soluble and -insoluble proteins and analyzed by Western blotting. The HSPB expression levels are shown in Fig. S1A in the supplemental material. (D) Immunofluorescent staining of Flag-tagged PARK2 (green), HSPBs (red), and DAPI (blue) in HEK293 cells, transfected as in panel C. (E) Diagram showing the percentages of flag-tagged PARK2-expressing cells with aggregates (*, P < 0.05; **, P < 0.001; n > three independent samples, means ± the SEM). Cells were divided in different categories, as shown in Fig. S1B in the supplemental material. HSPB6 was added as a negative control. HMW, high molecular weight.

Article Snippet: Mouse monoclonal anti-PARK2 antibody was obtained from Cell Signaling Technology.

Techniques: Transfection, Immunofluorescence, Western Blot, Expressing, Staining, Negative Control, High Molecular Weight

Journal: Molecular and Cellular Biology

Article Title: HSPA1A-Independent Suppression of PARK2 C289G Protein Aggregation by Human Small Heat Shock Proteins

doi: 10.1128/MCB.00698-14

Figure Lengend Snippet: Colocalization of the HSPBs with PARK2 C289G aggregates. (A) Immunofluorescent staining of Flag-tagged PARK2 (green), HSPBs (red), and DAPI (blue) in HEK293 cells, transfected as for Fig. 1D.

Article Snippet: Mouse monoclonal anti-PARK2 antibody was obtained from Cell Signaling Technology.

Techniques: Staining, Transfection

Journal: Molecular and Cellular Biology

Article Title: HSPA1A-Independent Suppression of PARK2 C289G Protein Aggregation by Human Small Heat Shock Proteins

doi: 10.1128/MCB.00698-14

Figure Lengend Snippet: HSPB1, HSPB2, HSPB4, and HSPB7 suppress PARK2 C289G aggregation in an HSPA1A-independent manner. (A) HEK293 cells were transfected with nonsense or HSPA1A RNAi. At 48 h posttransfection, the cells were transfected with Flag-tagged PARK2 C289G and mRFP-, HSPB1-, HSPB2-, HSPB4-, or HSPB7-encoding vectors and fractionated 24 h later in TX-100-soluble and -insoluble proteins. HMW, high molecular weight. (B) HEK293 cells were transfected for 24 h with Flag-tagged PARK2 C289G and mRFP-, HSPB1-, HSPB2-, HSPB4-, or HSPB7-encoding vector. Cells were treated with VER-155008 (40 μM, 24 h) and fractionated in TX-100-soluble and -insoluble proteins.

Article Snippet: Mouse monoclonal anti-PARK2 antibody was obtained from Cell Signaling Technology.

Techniques: Transfection, High Molecular Weight, Plasmid Preparation

Journal: Molecular and Cellular Biology

Article Title: HSPA1A-Independent Suppression of PARK2 C289G Protein Aggregation by Human Small Heat Shock Proteins

doi: 10.1128/MCB.00698-14

Figure Lengend Snippet: Role of ubiquitin proteasome system or the autophagy pathway in HSPB effects on PARK2 C289G aggregates. (A) HEK293 cells, transfected as in Fig. 3A, were treated with bortezomib (100 nM) or with 3-MA (20 mM) and wortmannin (200 nM) overnight. Cells were fractionated in TX-100 and analyzed for PARK2 C298G aggregates (*, P < 0.05; **, P < 0.001; n > three independent samples, means ± the SEM). (B) ATG5−/− MEF cells were transfected for 24 h with Flag-tagged PARK2 C289G and mRFP-, HSPB1-, HSPB2-, HSPB4-, or HSPB7-encoding vector and analyzed for PARK2 C298G aggregates (*, P < 0.05; **, P < 0.001; n > three independent samples, means ± the SEM). HMW, high molecular weight.

Article Snippet: Mouse monoclonal anti-PARK2 antibody was obtained from Cell Signaling Technology.

Techniques: Ubiquitin Proteomics, Transfection, Plasmid Preparation, High Molecular Weight