sdc1 polyclonal  (Proteintech)

Journal: Journal of Hematology & Oncology

Article Title: Fibroblast growth factor-2 (FGF2) and syndecan-1 (SDC1) are potential biomarkers for putative circulating CD15+/CD30+ cells in poor outcome Hodgkin lymphoma patients

doi: 10.1186/1756-8722-6-62

Figure Lengend Snippet: HL-relevant genes identified by bioinformatics data mining

Article Snippet: The antibodies used were the same as for IHC, except that a SDC1 rabbit polyclonal antibody (Sigma-Aldrich) was used for CD30-SDC1 double staining.

Techniques:

Journal: Journal of Hematology & Oncology

Article Title: Fibroblast growth factor-2 (FGF2) and syndecan-1 (SDC1) are potential biomarkers for putative circulating CD15+/CD30+ cells in poor outcome Hodgkin lymphoma patients

doi: 10.1186/1756-8722-6-62

Figure Lengend Snippet: FGF2 and SDC1 are overexpressed by HL cell lines and by CD30+ cells in the poor outcome HL patient group. (A) FGF2 and SDC1 expression in 10 different HL cell lines (solid black bar) is represented as the normalized fold change relative to purified normal B-cells (NBC, solid gray bar). The standard error (SE) for each cell line is indicated above each bar. See Table for HL cell line characteristics. (B) Qualitative mean intensity scores for FGF2 (solid black bar) and SDC1 (solid gray bar) from immunostained tissues in an array format consisting of 10 normal, 30 classical HL (cHL), and 18 Lymphocyte Predominant-HL (LP-HL) and 116 Non-HL (NHL) samples (y-axis). Immunostaining intensity was scored as 0 (no staining), 1 (weak), 2 (moderate), or 3 (intense). Standard error bars of the mean are indicated. (C) FGF2, SDC1, and CD30 mRNA expression levels in normal lymph node controls (NC, solid gray bar) and HL tissues associated with good outcome (GO, striped bar) and poor outcome (PO, solid black bar) were analyzed by qRT-PCR. The measurements represent the fold change after normalization with the NC group. (D) The same set of normal and HL tissues from (B) were immunostained for FGF2, SDC1, and CD30. Representative normal and stage II GO and PO patients are shown. (E) CD20 expression in normal lymph nodes and HL tissues analyzed by immunostaining. The significance of all qRT-PCR data comparing GO and PO is indicated (p<0.005). Scale bars represent 100 μm.

Article Snippet: The antibodies used were the same as for IHC, except that a SDC1 rabbit polyclonal antibody (Sigma-Aldrich) was used for CD30-SDC1 double staining.

Techniques: Expressing, Purification, Immunostaining, Staining, Quantitative RT-PCR

Journal: Journal of Hematology & Oncology

Article Title: Fibroblast growth factor-2 (FGF2) and syndecan-1 (SDC1) are potential biomarkers for putative circulating CD15+/CD30+ cells in poor outcome Hodgkin lymphoma patients

doi: 10.1186/1756-8722-6-62

Figure Lengend Snippet: CD30+ cells coexpress FGF2 and SDC1 in macrophage-rich HL tissues with poor outcome. (A) Double immunofluorescent staining showing expression of either FGF2 or SDC1 by CD30+ cells of poor outcome samples. Individual green or red fluorescence is depicted at the bottom of each image; scale bar (white solid bar) represents 100 μm. (B) Distribution of the immunophenotypes by outcome. The mean intensity scores for FGF2 (solid gray bar) and SDC1 (solid black bar) (y-axis) for the good outcome (GO) and poor outcome (PO) groups of HL patients. Immunofluorescence intensity was scored as 0 (no staining), 1 (weak), 2 (moderate), or 3 (strong) for FGF2+ or FGF2- and SDC1+ or SDC1-. The frequency (%) of expression of each combination of FGF2+/− and SDC1+/− among all tissue sections is indicated above each bar. (C) CD68 macrophage marker expression was analyzed by immunostaining (image) and qRT-PCR (graph) in normal lymph node control (NC), good outcome (GO), and poor outcome (PO) groups of HL patients. The fold-change in CD68 mRNA was calculated after normalization with NC. Significance of all qRT-PCR data comparing GO and PO is indicated for (B) and (C) (p < 0.005). Scale bars represent 100 μm.

Article Snippet: The antibodies used were the same as for IHC, except that a SDC1 rabbit polyclonal antibody (Sigma-Aldrich) was used for CD30-SDC1 double staining.

Techniques: Staining, Expressing, Fluorescence, Immunofluorescence, Marker, Immunostaining, Quantitative RT-PCR

Journal: Journal of Hematology & Oncology

Article Title: Fibroblast growth factor-2 (FGF2) and syndecan-1 (SDC1) are potential biomarkers for putative circulating CD15+/CD30+ cells in poor outcome Hodgkin lymphoma patients

doi: 10.1186/1756-8722-6-62

Figure Lengend Snippet: Metastatic markers TGFβ1 and MMP9 are overexpressed in poor outcome HL patients and by HL cell lines. (A) Protein and mRNA expression levels of TGFβ1 and MMP9 in normal lymph node control (NC), good outcome (GO) group and poor outcome (PO) group analyzed by immunostaining (left, images only for PO group) and qRT-PCR (right). mRNA expression is represented by fold-change (y-axis) after normalization with the control (NC). Significance of all qRT-PCR data comparing GO and PO is indicated (p < 0.005). TGFβ1 and MMP9 are also overexpressed by the HL cell lines (lower image of gel electrophoresis of (A) ). (B) TGFβ1 and MMP9 protein coexpression in tissues from the poor outcome HL patient group analyzed by double immunofluorescence staining for CD30, TGFβ1 and MMP9, or SDC1, TGFβ1 and MMP9. Individual green or red fluorescence is depicted at the bottom of each image. (C) Coexpression of TGFβ1 and MMP9 by subsets of tumor cells in poor outcome sample. (Inset of A and B ) Hodgkin Reed Sternberg cells (HRS) coexpressing SDC1 and TGFβ1 or SDC1 and MMP9. Scale bar (white solid bar) represents 100 μm.

Article Snippet: The antibodies used were the same as for IHC, except that a SDC1 rabbit polyclonal antibody (Sigma-Aldrich) was used for CD30-SDC1 double staining.

Techniques: Expressing, Immunostaining, Quantitative RT-PCR, Nucleic Acid Electrophoresis, Double Immunofluorescence Staining, Fluorescence

Journal: Journal of Hematology & Oncology

Article Title: Fibroblast growth factor-2 (FGF2) and syndecan-1 (SDC1) are potential biomarkers for putative circulating CD15+/CD30+ cells in poor outcome Hodgkin lymphoma patients

doi: 10.1186/1756-8722-6-62

Figure Lengend Snippet: FGF2 and SDC1 are overexpressed in circulating CD15+/CD30+ cells from chemo-naive poor outcome HL patients. qRT-PCR analysis of cells isolated from the buffy-coat of peripheral blood from normal donor controls (NC, striped bar), chemo- naïve (CN) good outcome (GO, dotted) and CN poor outcome (PO, solid black bar) groups, and chemo-exposed PO group (CE, checkered bar). Expression levels are represented as fold-change (y-axis) after normalization with normal control cells (N, solid gray bar: N denotes B cells in A and C; N denotes monocytes, CD4 T cells, CD8 T cells, and CD19 B cells in B). (A) mRNA expression of CD30 and CD15; (B) cell-specific markers for monocytes (CD14, CD63), T-cells (CD4,CD8), and B-cells (CD38, CD19); (C) FGF2 and SDC1. Significance of all qRT-PCR data comparing chemo-naïve GO and chemo-naïve PO is indicated (p < 0.0001; ANOVA and PLSD ).

Article Snippet: The antibodies used were the same as for IHC, except that a SDC1 rabbit polyclonal antibody (Sigma-Aldrich) was used for CD30-SDC1 double staining.

Techniques: Quantitative RT-PCR, Isolation, Expressing

Journal: Journal of Hematology & Oncology

Article Title: Fibroblast growth factor-2 (FGF2) and syndecan-1 (SDC1) are potential biomarkers for putative circulating CD15+/CD30+ cells in poor outcome Hodgkin lymphoma patients

doi: 10.1186/1756-8722-6-62

Figure Lengend Snippet: Primer sets for each gene used in this study

Article Snippet: The antibodies used were the same as for IHC, except that a SDC1 rabbit polyclonal antibody (Sigma-Aldrich) was used for CD30-SDC1 double staining.

Techniques: Sequencing

Journal: Biomolecules

Article Title: Mapping the Interactome of the Nuclear Heparan Sulfate Proteoglycan Syndecan-1 in Mesothelioma Cells

doi: 10.3390/biom10071034

Figure Lengend Snippet: Venn diagram representing overlap between SDC1 co-precipitating proteins and control sample (precipitate obtained with mouse IgG but without the addition of the CD138 antibody).

Article Snippet: The supernatants were collected followed by incubation overnight at 4 °C with corresponding Dynabead Protein A/G mixtures, which had been pre-incubated with polyclonal rabbit anti-SDC1 (Catalog no. 36-2900; Invitrogen, Gothenburg, Sweden; 100 µg per mL of mixture) and, as negative controls, beads were incubated with mouse IgG1 (Catalog no. X0931, Dako, CA, USA).

Techniques:

Journal: Biomolecules

Article Title: Mapping the Interactome of the Nuclear Heparan Sulfate Proteoglycan Syndecan-1 in Mesothelioma Cells

doi: 10.3390/biom10071034

Figure Lengend Snippet: The sub-cellular localization of proteins interacting with SDC1, classified into cytosol, mitochondria, nucleus and extracellular matrices.

Article Snippet: The supernatants were collected followed by incubation overnight at 4 °C with corresponding Dynabead Protein A/G mixtures, which had been pre-incubated with polyclonal rabbit anti-SDC1 (Catalog no. 36-2900; Invitrogen, Gothenburg, Sweden; 100 µg per mL of mixture) and, as negative controls, beads were incubated with mouse IgG1 (Catalog no. X0931, Dako, CA, USA).

Techniques:

Journal: Biomolecules

Article Title: Mapping the Interactome of the Nuclear Heparan Sulfate Proteoglycan Syndecan-1 in Mesothelioma Cells

doi: 10.3390/biom10071034

Figure Lengend Snippet: Significant pathways where SDC1 interacts with nuclear proteins.

Article Snippet: The supernatants were collected followed by incubation overnight at 4 °C with corresponding Dynabead Protein A/G mixtures, which had been pre-incubated with polyclonal rabbit anti-SDC1 (Catalog no. 36-2900; Invitrogen, Gothenburg, Sweden; 100 µg per mL of mixture) and, as negative controls, beads were incubated with mouse IgG1 (Catalog no. X0931, Dako, CA, USA).

Techniques: Infection, Non-Homologous End Joining

Journal: Biomolecules

Article Title: Mapping the Interactome of the Nuclear Heparan Sulfate Proteoglycan Syndecan-1 in Mesothelioma Cells

doi: 10.3390/biom10071034

Figure Lengend Snippet: The table represents interacting proteins of SDC1 having higher peak area values. The values were rounded off to the nearest whole number.

Article Snippet: The supernatants were collected followed by incubation overnight at 4 °C with corresponding Dynabead Protein A/G mixtures, which had been pre-incubated with polyclonal rabbit anti-SDC1 (Catalog no. 36-2900; Invitrogen, Gothenburg, Sweden; 100 µg per mL of mixture) and, as negative controls, beads were incubated with mouse IgG1 (Catalog no. X0931, Dako, CA, USA).

Techniques: RNA Binding Assay, Expressing

Journal: Biomolecules

Article Title: Mapping the Interactome of the Nuclear Heparan Sulfate Proteoglycan Syndecan-1 in Mesothelioma Cells

doi: 10.3390/biom10071034

Figure Lengend Snippet: Validation of interaction between SDC1 and EWSR1 or FUS, respectively. Western blots of SDC1 immunoprecipitates. ( a ) The blot shows presence of EWSR1. ( b ) The presence of FUS in SDC1 immunoprecipitate. “L” represents ladder in both the blots. ( c , d ) The figure shows the presence of SDC1 (red) along with EWSR1 (green) and FUS (green) in the nucleus, respectively.

Article Snippet: The supernatants were collected followed by incubation overnight at 4 °C with corresponding Dynabead Protein A/G mixtures, which had been pre-incubated with polyclonal rabbit anti-SDC1 (Catalog no. 36-2900; Invitrogen, Gothenburg, Sweden; 100 µg per mL of mixture) and, as negative controls, beads were incubated with mouse IgG1 (Catalog no. X0931, Dako, CA, USA).

Techniques: Western Blot

Journal: Biomolecules

Article Title: Mapping the Interactome of the Nuclear Heparan Sulfate Proteoglycan Syndecan-1 in Mesothelioma Cells

doi: 10.3390/biom10071034

Figure Lengend Snippet: Effect of loss- and gain-of-SDC1 on cellular amounts of total RNA and specific RNA for actin, relative to DNA content. ( a – c ; left images) Bar graphs showing the effects of knocking down SDC1 by siRNA (siSDC1) versus control (sictrl) on total RNA ( a ), actin ( b ), and GAPDH ( c ) RNA levels. ( a – c ; right images) Bar graphs showing the effects of overexpressing full-length SDC1 (FL) versus an empty vector (EV) on total RNA ( a ) actin ( b ) and GAPDH ( c ) RNA levels. The results are based on three independent experiments. Asterisks indicate statistical significance from the corresponding controls (* p < 0.05).

Article Snippet: The supernatants were collected followed by incubation overnight at 4 °C with corresponding Dynabead Protein A/G mixtures, which had been pre-incubated with polyclonal rabbit anti-SDC1 (Catalog no. 36-2900; Invitrogen, Gothenburg, Sweden; 100 µg per mL of mixture) and, as negative controls, beads were incubated with mouse IgG1 (Catalog no. X0931, Dako, CA, USA).

Techniques: Plasmid Preparation