NDRG2 Antibody (6A5) - Azide and BSA Free

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03]

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Western Blot: NDRG2 Antibody (6A5) [H00057447-M03]

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Western Blot: NDRG2 Antibody (6A5) [H00057447-M03]

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Western Blot: NDRG2 Antibody (6A5) [H00057447-M03]

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Western Blot: NDRG2 Antibody (6A5) [H00057447-M03]

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Immunocytochemistry/ Immunofluorescence: NDRG2 Antibody (6A5) [H00057447-M03]

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Western Blot: NDRG2 Antibody (6A5) [H00057447-M03]

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] - Analysis of NDRG2 expression in Hela S3 NE.

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03]

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Western Blot: NDRG2 Antibody (6A5) [H00057447-M03]

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ELISA: NDRG2 Antibody (6A5) [H00057447-M03]

ELISA: NDRG2 Antibody (6A5) [H00057447-M03] - Detection limit for recombinant GST tagged NDRG2 is approximately 0.03ng/ml as a capture antibody.

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] -

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] - NDRG2 inhibits cell proliferation & reduces intracellular glucose levels of breast cancer cells. (E) & (F) SK-BR-3 cells cultured to glucose medium at concentrations of 0, 5, 25, 50 & 100 mM for 24 hrs, & then the protein or mRNA was extracted for analysis by immunoblotting (E) or real-time PCR (F). beta -actin used as a loading control. The data presented means ± SD; error bars represented SD from 3 replicative wells. *P < 0.05 & **P < 0.01 versus control group. Image collected & cropped by CiteAb from the following publication (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] -

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] - NDRG2 inhibits cell proliferation & reduces intracellular glucose levels of breast cancer cells. (A) T-47D, MCF-7, Bcap37, MDA-MB-231 & SK-BR-3 cells collected for the extraction of proteins & analysed for N-myc downstream-regulated gene 2 (NDRG2) expression by immunoblotting. Image collected & cropped by CiteAb from the following publication (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] -

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] - NDRG2 interacts with GLUT1. (A) SK-BR-3 cells were fixed & incubated with primary antibodies against N-myc downstream-regulated gene 2 (NDRG2) or glucose transporter 1 (GLUT1) & with fluorescein isothiocyanate or a cyanine 3 secondary antibody. Green fluorescence indicates NDRG2 expression, red fluorescence indicates GLUT1 expression & blue fluorescence indicates nuclear staining. The results of the merged images reveal that NDRG2 & GLUT1 were colocalised in the cytoplasm. (B) Immunoprecipitation (IP) assays were performed with whole-cell lysates of SK-BR-3 cells pretreated with protein A–conjugated sepharose beads. Whole-cell lysates were probed for input. The antibodies for immunoprecipitation & Western blot (WB) analyses were carried out as indicated. The locations of various proteins are indicated by arrowheads. IgG, Immunoglobulin G. Image collected & cropped by CiteAb from the following publication (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: NDRG2 Antibody (6A5) [H00057447-M03] -

Immunocytochemistry/ Immunofluorescence: NDRG2 Antibody (6A5) [H00057447-M03] - NDRG2 decreases the glucose uptake & GLUT1 protein levels in SK-BR-3-based subcutaneously xenograft tumours. The experiments illustrated are described in the Methods section. (A) Tumour growth was assessed every 3 days until day 21 treatment by measuring two perpendicular diameters & calculating the volume in cubic centimetres. Ad-LacZ, adenovirus expressing LacZ; Ad-NDRG2, adenovirus expressing NDRG2; PFU, Plaque-forming units. The data presented are means ± SD; error bars represent SD from 6 mice. *P < 0.05 & **P < 0.01 versus phosphate-buffered saline (PBS) or Ad-LacZ. (B) Tumour cells were dissociated from xenograft tumours & suspended in PBS after the number of cells was counted. Next, the glucose uptake of cells in each group was detected. The data presented are means ± SD of three independent experiments; error bars represent SD from 6 mice. *P < 0.05 & **P < 0.01 versus PBS or Ad-LacZ. (C) Intratumoural protein expression was assessed by N-myc downstream-regulated gene 2 (NDRG2) & glucose transporter 1 (GLUT1) IHC staining. Representative images are shown. Original magnification: 400 x; Scale bars = 50 μm. (D) Proteins of the xenograft tumours from each group were extracted & analysed by immunoblotting to quantify NDRG2 & GLUT1 protein changes. Image collected & cropped by CiteAb from the following publication (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry: NDRG2 Antibody (6A5) [H00057447-M03] -

Immunohistochemistry: NDRG2 Antibody (6A5) [H00057447-M03] - NDRG2 is correlated with increased survival & negatively correlated with GLUT1 in breast carcinoma. Kaplan–Meier analysis was carried out according to N-myc downstream-regulated gene 2 (NDRG2) expression levels of disease-free survival (A) & overall survival (B). (C) Serial immunostained sections for NDRG2 & glucose transporter 1 (GLUT1) in breast cancer & normal tissues were analysed. Original magnification, 40× (top) & 400× (bottom); scale bars = 50 μm. (D) Protein was extracted from matched breast tumour tissue (T) & adjacent normal tissue (N) & subjected to immunoblot analysis to examine NDRG2 & GLUT1 expression. beta -actin served as a loading control. P: patient. Relative expression levels of NDRG2 (E) & GLUT1 (F) in human breast cancer & adjacent normal tissue are shown. immunoreactivity score distribution of cancer & adjacent normal tissue were represented with black & brown closed circles, respectively. The horizontal lines presented are means; error bars represented SD from 30 samples. P < 0.01 was considered a statistically significant difference. Image collected & cropped by CiteAb from the following publication (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: NDRG2 Antibody (6A5) [H00057447-M03] -

Immunocytochemistry/ Immunofluorescence: NDRG2 Antibody (6A5) [H00057447-M03] - NDRG2 interacts with GLUT1. (A) SK-BR-3 cells were fixed & incubated with primary antibodies against N-myc downstream-regulated gene 2 (NDRG2) or glucose transporter 1 (GLUT1) & with fluorescein isothiocyanate or a cyanine 3 secondary antibody. Green fluorescence indicates NDRG2 expression, red fluorescence indicates GLUT1 expression & blue fluorescence indicates nuclear staining. The results of the merged images reveal that NDRG2 & GLUT1 were colocalised in the cytoplasm. (B) Immunoprecipitation (IP) assays were performed with whole-cell lysates of SK-BR-3 cells pretreated with protein A–conjugated sepharose beads. Whole-cell lysates were probed for input. The antibodies for immunoprecipitation & Western blot (WB) analyses were carried out as indicated. The locations of various proteins are indicated by arrowheads. IgG, Immunoglobulin G. Image collected & cropped by CiteAb from the following publication (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] -

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] - NDRG2 downregulates GLUT1 by promoting its ubiquitination. (A), (C) & (E) SK-BR-3 cells were infected with an adenovirus carrying N-myc downstream-regulated gene 2 (Ad-NDRG2) at 1, 5 & 10 multiplicity of infection (MOI) or Ad-LacZ for 48 hours. (B), (D) & (F) T-47D cells were transfected with NDRG2 small interfering RNA (siRNA) 10, 25 & 100 pmol or control siRNA for 48 hours. Next, cell proteins or mRNA were extracted & analysed by immunoblotting (A) & (B) or by real-time PCR (C) to (F). beta -actin was used as a loading control. (C) – (F) The data presented are the means ± SD of three independent experiments; error bars represent SD from 3 replicative wells. *P < 0.05 & **P < 0.01 versus control group. (G) SK-BR-3 cells were infected with 10 MOI Ad-NDRG2 or Ad-LacZ for 48 hours & then treated with 2 μM, 6 μM or 8 μM MG-132 for 4 hours. Next, the protein was extracted & analysed by immunoblotting. (H) Cell fractions were prepared from the SK-BR-3 cells infected with 10 MOI Ad-NDRG2 or Ad-LacZ for 48 hours, & the membrane & cytosolic fractions of endogenous glucose transporter 1 (GLUT1) protein were detected. Tubulin & beta -actin served as loading controls. (I) SK-BR-3 cells were transfected with hemagglutinin (HA)-ubiquitin plasmid for 6 hours & infected with Ad-NDRG2 or Ad-LacZ for another 48 hours. Subsequently, the cell lysates were collected & analysed by immunoprecipitation (IP) & immunoblotting with GLUT1 & HA antibodies. WB, Western blot. Image collected & cropped by CiteAb from the following publication (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] -

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] - NDRG2 downregulates GLUT1 by promoting its ubiquitination. (A), (C) & (E) SK-BR-3 cells were infected with an adenovirus carrying N-myc downstream-regulated gene 2 (Ad-NDRG2) at 1, 5 & 10 multiplicity of infection (MOI) or Ad-LacZ for 48 hours. (B), (D) & (F) T-47D cells were transfected with NDRG2 small interfering RNA (siRNA) 10, 25 & 100 pmol or control siRNA for 48 hours. Next, cell proteins or mRNA were extracted & analysed by immunoblotting (A) & (B) or by real-time PCR (C) to (F). beta -actin was used as a loading control. (C) – (F) The data presented are the means ± SD of three independent experiments; error bars represent SD from 3 replicative wells. *P < 0.05 & **P < 0.01 versus control group. (G) SK-BR-3 cells were infected with 10 MOI Ad-NDRG2 or Ad-LacZ for 48 hours & then treated with 2 μM, 6 μM or 8 μM MG-132 for 4 hours. Next, the protein was extracted & analysed by immunoblotting. (H) Cell fractions were prepared from the SK-BR-3 cells infected with 10 MOI Ad-NDRG2 or Ad-LacZ for 48 hours, & the membrane & cytosolic fractions of endogenous glucose transporter 1 (GLUT1) protein were detected. Tubulin & beta -actin served as loading controls. (I) SK-BR-3 cells were transfected with hemagglutinin (HA)-ubiquitin plasmid for 6 hours & infected with Ad-NDRG2 or Ad-LacZ for another 48 hours. Subsequently, the cell lysates were collected & analysed by immunoprecipitation (IP) & immunoblotting with GLUT1 & HA antibodies. WB, Western blot. Image collected & cropped by CiteAb from the following publication (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] -

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] - NDRG2 decreases the glucose uptake & GLUT1 protein levels in SK-BR-3-based subcutaneously xenograft tumours. The experiments illustrated are described in the Methods section. (A) Tumour growth was assessed every 3 days until day 21 treatment by measuring two perpendicular diameters & calculating the volume in cubic centimetres. Ad-LacZ, adenovirus expressing LacZ; Ad-NDRG2, adenovirus expressing NDRG2; PFU, Plaque-forming units. The data presented are means ± SD; error bars represent SD from 6 mice. *P < 0.05 & **P < 0.01 versus phosphate-buffered saline (PBS) or Ad-LacZ. (B) Tumour cells were dissociated from xenograft tumours & suspended in PBS after the number of cells was counted. Next, the glucose uptake of cells in each group was detected. The data presented are means ± SD of three independent experiments; error bars represent SD from 6 mice. *P < 0.05 & **P < 0.01 versus PBS or Ad-LacZ. (C) Intratumoural protein expression was assessed by N-myc downstream-regulated gene 2 (NDRG2) & glucose transporter 1 (GLUT1) IHC staining. Representative images are shown. Original magnification: 400 x; Scale bars = 50 μm. (D) Proteins of the xenograft tumours from each group were extracted & analysed by immunoblotting to quantify NDRG2 & GLUT1 protein changes. Image collected & cropped by CiteAb from the following publication (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] -

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] - NDRG2 inhibits cell proliferation & reduces intracellular glucose levels of breast cancer cells. (B) SK-BR-3 cells with low NDRG2 expression infected by an adenovirus carrying NDRG2 (Ad-NDRG2) or negative control LacZ (Ad-LacZ), & T-47D cells with high NDRG2 transfected with small interfering RNA targeting NDRG2 (NDRG2 siRNA) or negative control siRNA (Con siRNA). Thereafter proteins extracted from these cells & analysed by immunoblotting. beta -actin used as a loading control. Before being cultured in 25 mM high-glucose (H.G.) or 5.5 mM low-glucose (L.G.) medium, SK-BR-3 cells infected by Ad-NDRG2 (C) & T-47D cells transfected by NDRG2 siRNA (D). Cell proliferation was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay for 1 to 5 days.Image collected & cropped by CiteAb from the following publication (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] -

Western Blot: NDRG2 Antibody (6A5) [H00057447-M03] - NDRG2 is correlated with increased survival & negatively correlated with GLUT1 in breast carcinoma. Kaplan–Meier analysis was carried out according to N-myc downstream-regulated gene 2 (NDRG2) expression levels of disease-free survival (A) & overall survival (B). (C) Serial immunostained sections for NDRG2 & glucose transporter 1 (GLUT1) in breast cancer & normal tissues were analysed. Original magnification, 40× (top) & 400× (bottom); scale bars = 50 μm. (D) Protein was extracted from matched breast tumour tissue (T) & adjacent normal tissue (N) & subjected to immunoblot analysis to examine NDRG2 & GLUT1 expression. beta -actin served as a loading control. P: patient. Relative expression levels of NDRG2 (E) & GLUT1 (F) in human breast cancer & adjacent normal tissue are shown. immunoreactivity score distribution of cancer & adjacent normal tissue were represented with black & brown closed circles, respectively. The horizontal lines presented are means; error bars represented SD from 30 samples. P < 0.01 was considered a statistically significant difference. Image collected & cropped by CiteAb from the following publication (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3628), licensed under a CC-BY license. Not internally tested by Novus Biologicals.