Journal: RNA

Article Title: A high-throughput pipeline for the production of synthetic antibodies for analysis of ribonucleoprotein complexes

doi: 10.1261/rna.055186.115

Figure Lengend Snippet: Validation of synthetic antibodies in the immunoprecipitation of endogenous target proteins. ( A ) One synthetic Fab against each of eight different RNP complex proteins, produced by the high-throughput pipeline, and ( B ) synthetic Fabs produced against HOW, FMR1, and ORB2 by low-throughput approaches, were used to carry out IPs from Drosophila embryo extract collected 0- to 3-h post-egg-laying. The success of the IPs was assessed by Western blots probed with conventional polyclonal or monoclonal antibodies against the same proteins. The percentage of the IP input loaded is indicated, and comparison of the IP input with the amount of protein present in the IP provides an estimate of the efficiency of the IP for each antibody. In all cases, negative control IPs were carried out with the control Fab C1 . White space between lanes indicates where lanes from the same blot were reordered.

Article Snippet: To determine the efficacy of the IPs, IP samples were run on Western blots, which were probed with conventional polyclonal or monoclonal antibodies that had been previously produced against the RNP complex proteins of interest, as follows: rabbit anti-NOS , rabbit anti-IMP , rabbit anti-MEI-P26 , rabbit anti-eIF4G , rabbit anti-EGL , rabbit anti-ME31B , rabbit anti-PABP , guinea pig anti-DP1 , rabbit anti-HOW (provided by Talila Volk), mouse anti-FMR1 5A11 , and mouse anti-ORB2 4G8 ( ); the anti-FMR1 and anti-ORB2 antibodies were obtained from the Developmental Studies Hybridoma Bank at The University of Iowa.

Techniques: Immunoprecipitation, Produced, High Throughput Screening Assay, Western Blot, Negative Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Inflammatory cytokines break down intrinsic immunological tolerance of human primary keratinocytes to cytosolic DNA.

doi: 10.4049/jimmunol.1302120

Figure Lengend Snippet: FIGURE 4. Assembly of the DNA-activated signalsome in kerati- nocytes is supported by cytokine treatment. (A) Human primary ker- atinocytes were treated with FITC-dsDNA (delivered with Lipofectamine [Lipo], 2 mg/ml), LL37 (5 mg/ml), or TNF-a (50 ng/ml), as indicated. The cells were fixed after 2 h, stained for IFI16, STING, and TBK1, and analyzed by confocal microscopy. White box indicates area displayed in zoom column. Scale bar, 10 mm. (B) The cells were treated as in (A), but fixed after 6 h of treatment for subsequent staining and analysis. Similar results were obtained in at least three independent experiments. (C) Ker- atinocytes were left untreated (UT) or stimulated with 50 ng/ml of TNF-a or IL-1b for 2 h. Cytoplasmic extracts were analyzed for IFI16 and b-actin by Western blotting.

Article Snippet: For Western blotting, the following specific Abs were used: IFI16 (1G7; Santa Cruz Biotechnology, Santa Cruz, CA), cGAS (C6orf150; Sigma-Aldrich), RCC1 (sc-1162; Santa Cruz Biotechnology), and b-actin (AC-15 HRP; Abcam). at M ichigan State U niversity on February 8, 2015 http://w w w .jim m unol.org/ D ow nloaded from

Techniques: Staining, Confocal Microscopy, Western Blot

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Inflammatory cytokines break down intrinsic immunological tolerance of human primary keratinocytes to cytosolic DNA.

doi: 10.4049/jimmunol.1302120

Figure Lengend Snippet: FIGURE 5. DNA-driven gene expression in cyto- kine-treated keratinocytes is dependent on IFI16. (A) Human primary keratinocytes were transfected with control (Ctrl) and IFI16-specific siRNA. Total RNA was harvested 48 h later, and levels of IFI16 mRNA were measured by RT-qPCR. (B and C) Keratinocytes treated with siRNA for 48 h were stimulated with synthetic dsDNA (Lipofectamine [Lipo], 2 mg/ml) or TNF-a (50 ng/ml) as indicated for 6 h. Total RNAwas isolated, and mRNA levels of CCL20 and CXCL10 were quantified by RT-qPCR. The data were normal- ized to b-actin and are presented as mean fold in- duction of triplicate cultures 6 SD. Similar results were obtained in at least three independent experi- ments. *p , 0.05. NR, Normalized ratio.

Article Snippet: For Western blotting, the following specific Abs were used: IFI16 (1G7; Santa Cruz Biotechnology, Santa Cruz, CA), cGAS (C6orf150; Sigma-Aldrich), RCC1 (sc-1162; Santa Cruz Biotechnology), and b-actin (AC-15 HRP; Abcam). at M ichigan State U niversity on February 8, 2015 http://w w w .jim m unol.org/ D ow nloaded from

Techniques: Gene Expression, Transfection, Control, Quantitative RT-PCR, Isolation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Inflammatory cytokines break down intrinsic immunological tolerance of human primary keratinocytes to cytosolic DNA.

doi: 10.4049/jimmunol.1302120

Figure Lengend Snippet: FIGURE 6. IFI16 is upregulated in psoriasis lesions and exhibits a cellular localization pattern similar to keratinocytes receiving dual stimulation with DNA and inflammatory cytokines. (A) Tissue sections of punch biopsies from nonlesional and lesional psoriatic skin were stained for IFI16 and DNA (DAPI) and analyzed by confocal microscopy. Arrows indicate cytosolic IFI16 foci. Scale bar, 100 mm. (B) RNA was isolated from nonlesional (NLS) and lesional psoriatic skin (LS), and levels of IFI16 mRNA were quantified by RT- qPCR. Data shown as means of normalized values relative to nonlesional psoriasis skin 6 SD. n = 9. (C) Tissue section of punch biopsies from lesional psoriatic skin stained for DNA (DAPI) and analyzed by confocal microscopy. Arrows indicate DAPI staining (DNA) in the cytosol. Scale bar, 10 mm. *p , 0.05. DIC, Dif- ferential interference contrast; NR, Normalized ratio.

Article Snippet: For Western blotting, the following specific Abs were used: IFI16 (1G7; Santa Cruz Biotechnology, Santa Cruz, CA), cGAS (C6orf150; Sigma-Aldrich), RCC1 (sc-1162; Santa Cruz Biotechnology), and b-actin (AC-15 HRP; Abcam). at M ichigan State U niversity on February 8, 2015 http://w w w .jim m unol.org/ D ow nloaded from

Techniques: Staining, Confocal Microscopy, Isolation, Quantitative RT-PCR

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Immunohistochemical and Biochemical Expression Patterns of TTF-1, RAGE, GLUT-1 and SOX2 in HCV-Associated Hepatocellular Carcinomas

doi: 10.22034/APJCP.2018.19.1.219

Figure Lengend Snippet: Difference in Tissue Expression Scores of Studied Markers in Examined Lesions

Article Snippet: 4) SOX2 Antibody (NBP2-29623) (Novus Biologicals, USA) at an optimal dilution of 1:100 in PBS.

Techniques: Expressing

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Immunohistochemical and Biochemical Expression Patterns of TTF-1, RAGE, GLUT-1 and SOX2 in HCV-Associated Hepatocellular Carcinomas

doi: 10.22034/APJCP.2018.19.1.219

Figure Lengend Snippet: Difference in Markers Expression Scores in Different Ggrades of Hepatitis Activity and Stages of Fibrosis

Article Snippet: 4) SOX2 Antibody (NBP2-29623) (Novus Biologicals, USA) at an optimal dilution of 1:100 in PBS.

Techniques: Expressing, Activity Assay

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Immunohistochemical and Biochemical Expression Patterns of TTF-1, RAGE, GLUT-1 and SOX2 in HCV-Associated Hepatocellular Carcinomas

doi: 10.22034/APJCP.2018.19.1.219

Figure Lengend Snippet: Relation between the Expression of Tissue Markers and the Grades of HCC

Article Snippet: 4) SOX2 Antibody (NBP2-29623) (Novus Biologicals, USA) at an optimal dilution of 1:100 in PBS.

Techniques: Expressing

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Immunohistochemical and Biochemical Expression Patterns of TTF-1, RAGE, GLUT-1 and SOX2 in HCV-Associated Hepatocellular Carcinomas

doi: 10.22034/APJCP.2018.19.1.219

Figure Lengend Snippet: The Correlation between Serum TTF-1, RAGE, GLUT-1 and SOX-2 Genes Expression in Relation to Tumor Grade and Stage of Liver Fibrosis

Article Snippet: 4) SOX2 Antibody (NBP2-29623) (Novus Biologicals, USA) at an optimal dilution of 1:100 in PBS.

Techniques: Expressing

Journal: Asian Pacific Journal of Cancer Prevention : APJCP

Article Title: Immunohistochemical and Biochemical Expression Patterns of TTF-1, RAGE, GLUT-1 and SOX2 in HCV-Associated Hepatocellular Carcinomas

doi: 10.22034/APJCP.2018.19.1.219

Figure Lengend Snippet: The Validation of SOX-2, GLUT-1, RAGE and TTF-1 Genes as a Diagnostic Biomarker in the Serum

Article Snippet: 4) SOX2 Antibody (NBP2-29623) (Novus Biologicals, USA) at an optimal dilution of 1:100 in PBS.

Techniques: Biomarker Discovery, Diagnostic Assay

Journal: PLoS ONE

Article Title: Zileuton Improves Memory Deficits, Amyloid and Tau Pathology in a Mouse Model of Alzheimer’s Disease with Plaques and Tangles

doi: 10.1371/journal.pone.0070991

Figure Lengend Snippet: Antibodies used in this study.

Article Snippet: 4G8 , aa 18–22 of human beta amyloid (VFFAE) , Mouse , IHC , Covance.

Techniques: Transduction, Immunohistochemistry-IF, Recombinant

Journal: PLoS ONE

Article Title: Zileuton Improves Memory Deficits, Amyloid and Tau Pathology in a Mouse Model of Alzheimer’s Disease with Plaques and Tangles

doi: 10.1371/journal.pone.0070991

Figure Lengend Snippet: A, B. RIPA-soluble (RIPA) and formic acid extractable (FA) Aβ1-40 and Aβ1-42 levels in cortex of 3×Tg receiving zileuton or vehicle were measured by sandwich ELISA. (n = 9 for control, and n = 9 for zileuton; *p<0.05). C. Representative sections of brains from 3×Tg mice receiving zileuton or placebo (control) immunostained with 4G8 antibody. D. Quantification of the area occupied by Aβ immunoreactivity in brain of 3×Tg mice receiving zileuton or placebo (n = 3 control, and n = 3 zileuton; *p<0.05). E. Representative western blots of APP, ADAM-10, BACE-1, sAPPs, CTFs, PS1, Nicastrin, Pen-2, and APH-1 in the cortex of 3×Tg mice receiving zileuton or vehicle. F. Densitometric analyses of the immunoreactivities to the antibodies shown in the previous panel. (n = 4, *p<0.05). Values represent mean ± SEM.

Article Snippet: 4G8 , aa 18–22 of human beta amyloid (VFFAE) , Mouse , IHC , Covance.

Techniques: Sandwich ELISA, Control, Western Blot