mouse anti human sod1  (Proteintech)

Journal: Cancer gene therapy

Article Title: Targeting SOD1 via RNAi with PEGylated graphene oxide nanoparticles in platinum-resistant ovarian cancer.

doi: 10.1038/s41417-023-00659-2

Figure Lengend Snippet: Fig. 3 Nanoparticle behaviour in biological solutions. A Colloidal stability of GO, GOPEI, and GOPEI-mPEG were assessed in DIW, 0.9% NaCl, and RPMI + 10% FBS, 1 h after treatment. Nanoparticle aggregation was observed in all three biological solutions containing non- functionalized GO. B, C Gel-electrophoresis-based serum stability was carried out in GOPEI1x, GOPEI2x, and GOPEI3x. The susceptibility of siRNA to RNase degradation in serum was assessed for 48 h. Naked siSCR was used as a control. D Kinetic study of nanocarrier-siRNA serum stability was measured using mouse serum in siSCRFAM conjugated GOPEI, GOPEI-mPEG, and Lipo2000 relative to control naked siSCRFAM. The fluorescence of siSCRFAM was measured over a time of up to 8 h. E Kinetic study of nanoparticle behaviour was conducted in vitro using siSCRFAM complexed GO, GOPEI and GOPEI-mPEG. The degree of quenching induced by GOPEI upon binding siSCRFAM was measured by a Varioskan fluorescent plate reader. A 10 h time-course study of the live cell uptake of the nanoparticles was measured by recording their fluorescent emission. Samples containing only cells and RPMI media were used as negative controls. F Confocal laser scanning images showing the cellular uptake of GOPEI- siSCRFAM. Merged fluorescence images of cells treated with GOPEI- siSCRFAM with a magnified image of a single A2780 cell showing successful transfection of GOPEI as aggregations were compared with Lipo2000 as the positive control, and naked siSCRFAM was used as a negative control. G Quantitative assessment of nanoparticle: siRNA cellular uptake GOPEI-siRNA transfection efficiency was analyzed using flow cytometry. Two siSCRFAM concentrations of 60 nM and 90 nM complexed with Lipo2000 served as positive control. H Image showing intracellular graphene accumulation in cell pellets following three washing steps and centrifugation prior to protein isolation. I, J Time-course study showing SOD1 mRNA and protein levels following SOD1 knockdown. K, L The effect of concentration dependent SOD1 knockdown on protein levels was determined by Western blot and mRNA expression levels were measured using RT-qPCR. All values are expressed as mean ± SD. ns- not significant, *p < 0.05, **p < 0.01, and ****p < 0.0001 as analyzed using two-tailed unpaired t-test.

Article Snippet: The membrane was incubated with mouse anti-human SOD1 (15.9 kDa) monoclonal antibody (Cat# 67480-1-Ig, 1:1000, ProteinTech, Rosemont, IL, USA) and with mouse β-actin (42 kDa) monoclonal antibody (1:2000, Cat# 66009-1-Ig, ProteinTech) overnight at 40 °C.

Techniques: Nucleic Acid Electrophoresis, Control, In Vitro, Binding Assay, Transfection, Positive Control, Negative Control, Cytometry, Centrifugation, Isolation, Knockdown, Concentration Assay, Western Blot, Expressing, Quantitative RT-PCR, Two Tailed Test

Journal: Cancer gene therapy

Article Title: Targeting SOD1 via RNAi with PEGylated graphene oxide nanoparticles in platinum-resistant ovarian cancer.

doi: 10.1038/s41417-023-00659-2

Figure Lengend Snippet: Fig. 4 In vitro nanoparticle toxicity. A Schematic diagram of the in vitro induction of acquired platinum resistance. B Baseline cisplatin sensitivity of A2780 and A2780DDP cell lines. C The IC50 of A2780 and A2780DDP were 4.71 ± 0.26 and 13.95 ± 1.18 µg/ml, respectively. D Bar graph showing normalized baseline SOD1 mRNA levels (n = 6). E Western blot showing the SOD1 protein levels in A2780 and A2780DDP cell lines in vivo in Q1 and Q3. F Bar graph showing normalized baseline SOD1 mRNA (n = 3). G Schematic illustration of xenograft tumour sample preparation for immunoblotting and qRT-PCR. H Western blot showing the SOD1 protein levels in A2780 and A2780DDP cell-derived xenograft tumour tissue samples. I Cisplatin treatment of A2780DDP cells (n = 6) induced SOD1 mRNA overexpression in a concentration-dependent manner measured by RT-qPCR. J Time-course evaluation SOD1 mRNA induction by GOPEI treatment in A2780DDP cells (n = 6) measured by qRT-PCR. K Cytotoxicity of GO, PEI, GOPEI and GOPEI-mPEG were determined at the following concentrations: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 25, 30, 35 and 40 µg/mL, respectively. L MTT assay showing relative cell viability after A2780 cells were treated with 9 µg/mL of GOPEI for 48 h followed by cisplatin treatment at 14 different concentrations (2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26 and 28 µg/mL respectively). IC50 values were compared with that of A2780 cells (n = 6). M Volcano plot showing the transcriptional activation of the mitochondrial unfolded protein response by 15 µM cisplatin in A2780 cell line. N, O GOPEI and GOPEI-mPEG treatment-induced differential in vitro activation of the UPRmt in (N) A2780 and (O) A2780DDP cell lines. P Schematic diagram illustrating UPRmt activation by cisplatin, graphene and cationic polymers leading to mitochondrial dysfunction and subsequent mito-nuclear signalling.

Article Snippet: The membrane was incubated with mouse anti-human SOD1 (15.9 kDa) monoclonal antibody (Cat# 67480-1-Ig, 1:1000, ProteinTech, Rosemont, IL, USA) and with mouse β-actin (42 kDa) monoclonal antibody (1:2000, Cat# 66009-1-Ig, ProteinTech) overnight at 40 °C.

Techniques: In Vitro, Western Blot, In Vivo, Sample Prep, Quantitative RT-PCR, Derivative Assay, Over Expression, Concentration Assay, MTT Assay, Activation Assay

Journal: Frontiers in Oncology

Article Title: LCS-1 inhibition of superoxide dismutase 1 induces ROS-dependent death of glioma cells and degradates PARP and BRCA1

doi: 10.3389/fonc.2022.937444

Figure Lengend Snippet: The expression of SOD1 in glioma tissues. (A) SOD1 IHC scores of normal brain tissues, tumor adjacent tissues and glioma grade I, II, III and IV tissues. * P < 0.05 compared with the normal brain tissue (Nor) or glioma grade I tissues. (B) Staining intensity of glioma with different histopathological types. Staining intensity was scored using a four-tier scale and defined as follows: negative staining (0-80); weak staining (80-200); moderate staining (200-300); strong staining (300-400). Nor, normal brain tissue; Adj, glioma adjacent tissues; I, well differentiated glioma; II, moderately differentiated glioma; III, poorly differentiated glioma; and IV, non-differentiated glioma. (C) SOD1 IHC staining for full glioma microarray. (D-E) Representative SOD1 weak (D) , moderate staining (E) in normal brain tissues. (F-H) Representative SOD1 weak (F) , moderate (G) , strong staining (H) in glioma adjacent tissues. (I-K) Representative SOD1 weak (I) , moderate (J) , strong staining (K) in glioma grade I tissues. (L, M) Representative SOD1 weak (L) , moderate (M) , strong staining (N) in glioma grade II tissues. (O-Q) Representative SOD1 weak (O) , moderate (P) , strong staining (Q) in glioma grade III tissues. (R) Representative SOD1 strong staining in glioma grade IV tissues.

Article Snippet: Mouse monoclonal anti-human SOD1 (sc-101523), SOD2 (sc-137254), BRCA1 (sc-6954) and BRCA2 (sc-518154) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Expressing, Staining, Negative Staining, Immunohistochemistry, Microarray

Journal: Frontiers in Oncology

Article Title: LCS-1 inhibition of superoxide dismutase 1 induces ROS-dependent death of glioma cells and degradates PARP and BRCA1

doi: 10.3389/fonc.2022.937444

Figure Lengend Snippet: The expression of SOD1, SOD2 and SOD3 in glioma cell lines. (A) qRT-PCR analysis of SOD1, SOD2 and SOD3 mRNA levels in U251 cells. (B) qRT-PCR analysis of SOD1, SOD2 and SOD3 mRNA levels in U87 cells. (C) FACS analysis of SOD1 and SOD2 protein levels in U251 and U87 cells. (D) qRT-PCR analysis of SOD1 and 2 mRNA levels in U251 cells treated with 10 µM LCS-1 for the indicated time periods. * P < 0.05 compared with the medium groups. (E) qRT-PCR analysis of SOD1, SOD2 and SOD3 mRNA levels in U87 cells treated with 10 µM LCS-1 for the indicated time periods. * P < 0.05 compared with the medium groups. (F, G) Western blot analysis of the protein levels of SOD1 in U251 (F) and U87 (G) cells. (H, I) The quantitative data from F (H) and G (I) respectively. * P < 0.05 compared with the medium groups.

Article Snippet: Mouse monoclonal anti-human SOD1 (sc-101523), SOD2 (sc-137254), BRCA1 (sc-6954) and BRCA2 (sc-518154) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Oncology

Article Title: LCS-1 inhibition of superoxide dismutase 1 induces ROS-dependent death of glioma cells and degradates PARP and BRCA1

doi: 10.3389/fonc.2022.937444

Figure Lengend Snippet: Model of the mechanism by which LCS-1 induces cell death. LCS-1 inhibited the enzyme activity of SOD1, resulting in the accumulation of ROS, leading to the induction of DNA damage. Meanwhile, LCS-1 induces the degradation of PARP. The dysfunction of PARP inhibits DNA damage repair via blocking both PARylation-mediated and EJ-mediated pathways. Furthermore, LCS-1 induces the degradation of BRCA1, eliciting the block of HR-mediated pathway. The inhibition of these three repair pathways results in death of glioma cells.

Article Snippet: Mouse monoclonal anti-human SOD1 (sc-101523), SOD2 (sc-137254), BRCA1 (sc-6954) and BRCA2 (sc-518154) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Activity Assay, Blocking Assay, Inhibition

Journal: Brain Pathology

Article Title: Accumulation of misfolded SOD1 outlines distinct patterns of motor neuron pathology and death during disease progression in a SOD1 G93A mouse model of amyotrophic lateral sclerosis

doi: 10.1111/bpa.13078

Figure Lengend Snippet: Antibodies used for immunocytochemistry

Article Snippet: SOD1 (C4F6) Anti‐Misfolded Human SOD1 , Mouse monoclonal , MediMabs 2B Scientific (MM‐00070‐2‐P) , 1:100.

Techniques: Ubiquitin Proteomics