Review

ebm basal medium (Chem Impex International)

Chem Impex International is a verified supplier

Chem Impex International manufactures this product

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Chem Impex International ebm basal medium
Ebm Basal Medium, supplied by Chem Impex International, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/00479/Phenol+red/pmc05321203-45-12-5
Average 95 stars, based on 1 article reviews

ebm basal medium - by Bioz Stars, 2026-09

95/100 stars

Images

Cell Culture:

Article Title: Single Step Synthesis, Characterization and Applications of Curcumin Functionalized Iron Oxide Magnetic Nanoparticles
Article Snippet: Iron (III) chloride hexahydrate (FeCl 3 .6H 2 O), iron (II) chloride (FeCl 2 .4H 2 O) and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich, while the ammonium hydroxide (NH 4 OH) was procured from EMD Chemicals. .. Curcumin powder was purchased from Chem-Impex Int’l Inc. For cell culture studies, EBM basal medium (phenol red free), EGM-2 growth factors, and Human Umbilical Vein Endothelial Cells were purchased from Lonza. .. Calcein-AM red-orange was purchased from Life Technologies.

Image Search Results

Angiogenic factors are upregulated in Smad1/5 dKO granulosa cell tumors. Real-time quantitative PCR was performed to determine expression levels of Pdgfa (A), Pdgfb (B), Vegfa (C) and Vegfb (D) in wild type mouse granulosa cells (GC), Smad1/5 dKO tumors (TU) and metastasis (MET). siRNA-mediated knockdown (+KD) of both Smad1 and Smad5 in mouse granulosa cells resulted in more than 85% reduction in Smad1 (E) and Smad5 (F) transcript levels compared to the cells transfected cells with a scrambled control siRNA (-KD). A significant increase in Pdgfa (G) expression is observed following Smad1/5 knockdown, without any change in Pdgfb (H) levels. Different letters above the bars indicate statistically different means by ANOVA and post hoc analysis (n=4; P< 0.05).

Journal: Oncogene

Article Title: BMP- specific SMADs function as novel repressors of PDGFA and modulate its expression in ovarian granulosa cells and tumors

doi: 10.1038/onc.2012.392

Figure Lengend Snippet: Angiogenic factors are upregulated in Smad1/5 dKO granulosa cell tumors. Real-time quantitative PCR was performed to determine expression levels of Pdgfa (A), Pdgfb (B), Vegfa (C) and Vegfb (D) in wild type mouse granulosa cells (GC), Smad1/5 dKO tumors (TU) and metastasis (MET). siRNA-mediated knockdown (+KD) of both Smad1 and Smad5 in mouse granulosa cells resulted in more than 85% reduction in Smad1 (E) and Smad5 (F) transcript levels compared to the cells transfected cells with a scrambled control siRNA (-KD). A significant increase in Pdgfa (G) expression is observed following Smad1/5 knockdown, without any change in Pdgfb (H) levels. Different letters above the bars indicate statistically different means by ANOVA and post hoc analysis (n=4; P< 0.05).

Article Snippet: Following transfer and blocking in 5% nonfat dry milk in Tris-buffered saline containing 0.1% Tween-20 (TBST) for one hour, the membrane was then incubated overnight at 4 °C with rabbit anti-phospho-Smad1/5 (Ser463/465) antibody (Cell Signaling, 41D10) diluted at 1:1000 or mouse anti-human SMAD1 antibody (Biomatrix research, BMR 00479) diluted at 1:500 or rabbit anti-human PDGF-A antibody (Santa Cruz, N-30) diluted at 1:500 or mouse anti-FLAG M2 antibody (Sigma- Aldrich, A8592) diluted at 1:1000 or mouse anti-β-actin antibody (Sigma- Aldrich, A5316) diluted at 1:10,000.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Knockdown, Transfection, Control

Regulation of Pdgfa is independent of BMP signaling through type I receptor. Cells were the treated with vehicle control medium or with 100 ng/ml BMP7 in the presence or absence of 10 μM dorsomorphin (DM) for 5h and 12h. Cells were harvested for total RNA and protein for real-time PCR and Western blotting. DM markedly reduced Id1 induction by BMP7 at 5h (A) and 12h (D), while no change is observed in Pdgfa expression (B and E). (C and F) Phosphorylation of SMAD1/5/8 in mouse granulosa cells detected by immunoblot after pretreatment with DM for 30 min followed by treatment with BMP7 for 5h and 12h. The membrane was stripped and re-probed for β-actin antibody for loading control. Different letters above the bars indicate statistical significance (n=3; P< 0.05).

Journal: Oncogene

Article Title: BMP- specific SMADs function as novel repressors of PDGFA and modulate its expression in ovarian granulosa cells and tumors

doi: 10.1038/onc.2012.392

Figure Lengend Snippet: Regulation of Pdgfa is independent of BMP signaling through type I receptor. Cells were the treated with vehicle control medium or with 100 ng/ml BMP7 in the presence or absence of 10 μM dorsomorphin (DM) for 5h and 12h. Cells were harvested for total RNA and protein for real-time PCR and Western blotting. DM markedly reduced Id1 induction by BMP7 at 5h (A) and 12h (D), while no change is observed in Pdgfa expression (B and E). (C and F) Phosphorylation of SMAD1/5/8 in mouse granulosa cells detected by immunoblot after pretreatment with DM for 30 min followed by treatment with BMP7 for 5h and 12h. The membrane was stripped and re-probed for β-actin antibody for loading control. Different letters above the bars indicate statistical significance (n=3; P< 0.05).

Techniques: Control, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Phospho-proteomics, Membrane

Loss of SMAD 1/5 upregulates PDGFA in a human juvenile granulosa cell tumor-derived cell line (COV434). Real-time PCR was performed to compare the expression levels of SMAD1 (A) and SMAD5 (B), PDGFA (C) and PDGFB (D) in human embryonic kidney cell-line HEK293T (HEK) and human granulosa cell tumor derived cell line (COV434). Transfection of scrambled control siRNA (-KD) or siRNAs specific for SMAD1 and SMAD5 (+KD) resulted in a significant reduction of SMAD1 (E) and SMAD5 (F) in COV434 cells. Concomitantly, a significant 2-fold increase in PDGFA (G) is achieved following SMAD1 and SMAD5 reduction by siRNA, without a measurable change in PDGFB (H). Different letters above the bars indicate statistical significance (n=3; P< 0.05).

Journal: Oncogene

Article Title: BMP- specific SMADs function as novel repressors of PDGFA and modulate its expression in ovarian granulosa cells and tumors

doi: 10.1038/onc.2012.392

Figure Lengend Snippet: Loss of SMAD 1/5 upregulates PDGFA in a human juvenile granulosa cell tumor-derived cell line (COV434). Real-time PCR was performed to compare the expression levels of SMAD1 (A) and SMAD5 (B), PDGFA (C) and PDGFB (D) in human embryonic kidney cell-line HEK293T (HEK) and human granulosa cell tumor derived cell line (COV434). Transfection of scrambled control siRNA (-KD) or siRNAs specific for SMAD1 and SMAD5 (+KD) resulted in a significant reduction of SMAD1 (E) and SMAD5 (F) in COV434 cells. Concomitantly, a significant 2-fold increase in PDGFA (G) is achieved following SMAD1 and SMAD5 reduction by siRNA, without a measurable change in PDGFB (H). Different letters above the bars indicate statistical significance (n=3; P< 0.05).

Techniques: Derivative Assay, Real-time Polymerase Chain Reaction, Expressing, Transfection, Control

SMAD1/5 bind to the PDGFA promoter. (A) In silico promoter analysis identified several putative SMAD1/5 binding sites (gray shaded region) in the human PDGFA promoter region. Also shown are binding sites for Sp1 (dashed boxes), a known positive regulator for PDGFA promoter activity. The arrows in the promoter region indicate the sites where forward (-294F) and reverse (-82R) primers are designed for the ChIP assay. (B) Chromatin immunoprecipitation (ChIP) assay with an anti-SMAD1/5 antibody or control IgG was performed on cell extracts from COV434 cells demonstrated the in vivo binding of SMAD1/5 to the PDGFA promoter. (C) Real-time PCR analysis of immunoprecipitated DNA using locus specific primers demonstrated three-fold enrichment (ChIP/Input DNA) of SMAD1/5 occupancy at the PDGFA promoter compared to negative control (no locus-specific primers) and IgG. Asterisks indicate statistical significance (n=3; P< 0.05).

Journal: Oncogene

Article Title: BMP- specific SMADs function as novel repressors of PDGFA and modulate its expression in ovarian granulosa cells and tumors

doi: 10.1038/onc.2012.392

Figure Lengend Snippet: SMAD1/5 bind to the PDGFA promoter. (A) In silico promoter analysis identified several putative SMAD1/5 binding sites (gray shaded region) in the human PDGFA promoter region. Also shown are binding sites for Sp1 (dashed boxes), a known positive regulator for PDGFA promoter activity. The arrows in the promoter region indicate the sites where forward (-294F) and reverse (-82R) primers are designed for the ChIP assay. (B) Chromatin immunoprecipitation (ChIP) assay with an anti-SMAD1/5 antibody or control IgG was performed on cell extracts from COV434 cells demonstrated the in vivo binding of SMAD1/5 to the PDGFA promoter. (C) Real-time PCR analysis of immunoprecipitated DNA using locus specific primers demonstrated three-fold enrichment (ChIP/Input DNA) of SMAD1/5 occupancy at the PDGFA promoter compared to negative control (no locus-specific primers) and IgG. Asterisks indicate statistical significance (n=3; P< 0.05).

Techniques: In Silico, Binding Assay, Activity Assay, Chromatin Immunoprecipitation, Control, In Vivo, Real-time Polymerase Chain Reaction, Immunoprecipitation, Negative Control

Induction of the human PDGFA promoter following knockdown of SMAD1 and SMAD5 . Luciferase reporter plasmids containing -881 or -261 base pairs of the human PDGFA promoter were transfected into wild type mouse granulosa cells (A) and human COV434 cells (B) following transfection of control (scrambled) siRNA (-KD) or siRNAs specific for SMAD1 and SMAD5 (+KD) along with the renilla luciferase control plasmid (pRL-TK). Twenty-four hours after transfection, the cells were lysed and assessed for luciferase activity. Luciferase activity from both the constructs was significantly increased in the absence of SMAD1 and SMAD5 . Data are presented as the ratio of firefly luciferase to renilla. Different letters above the bars indicate statistically different means by ANOVA and post hoc analysis (n=4; P< 0.05). pGL3B indicates the pGL3-Basic promoterless parent plasmid.

Journal: Oncogene

Article Title: BMP- specific SMADs function as novel repressors of PDGFA and modulate its expression in ovarian granulosa cells and tumors

doi: 10.1038/onc.2012.392

Figure Lengend Snippet: Induction of the human PDGFA promoter following knockdown of SMAD1 and SMAD5 . Luciferase reporter plasmids containing -881 or -261 base pairs of the human PDGFA promoter were transfected into wild type mouse granulosa cells (A) and human COV434 cells (B) following transfection of control (scrambled) siRNA (-KD) or siRNAs specific for SMAD1 and SMAD5 (+KD) along with the renilla luciferase control plasmid (pRL-TK). Twenty-four hours after transfection, the cells were lysed and assessed for luciferase activity. Luciferase activity from both the constructs was significantly increased in the absence of SMAD1 and SMAD5 . Data are presented as the ratio of firefly luciferase to renilla. Different letters above the bars indicate statistically different means by ANOVA and post hoc analysis (n=4; P< 0.05). pGL3B indicates the pGL3-Basic promoterless parent plasmid.

Techniques: Knockdown, Luciferase, Transfection, Control, Plasmid Preparation, Activity Assay, Construct

SMAD1/5 expression antagonizes Sp1 induction of PDGFA promoter. (A) COV434 cells were transiently co-transfected with 1 μg of -881:luc of the human PDGFA promoter and 0.5 μg of Sp1 and SMAD1/5 along with the renilla luciferase control plasmid (pRL-TK). Forty-eight hours after transfection, the cells were lysed and assessed for luciferase activity. Data are presented as the ratio of firefly luciferase to renilla. (B) Representative Western blot of whole cell lysates from COV434 cells transfected with either pcDNA3.1 or Flag-tagged SMAD1/5 expression plasmids and blotted with mouse anti-FLAG M2 antibody and mouse anti-β-actin antibody (loading control). (C) ChIP analysis in COV434 cells transfected with pcDNA3.1 or Flag-tagged SMAD1/5 expression plasmids. Chromatin cross-linked protein DNA complexes were immunoprecipitated with either anti-Sp1 antibody or with non-specific IgG and the PDGF-A promoter amplified by real-time PCR using locus specific primers. (D) SMAD1/5 represses the wild-type PDGFA promoter (-261:luc) but not the promoter bearing a mutation in the SMAD1/5 binding site (-261mutA: luc). COV434 cells were transiently co-transfected with 1 μg of -261:luc (control) or -261mutA: luc (mutant) and 0.5 μg of Sp1 and SMAD1/5 along with the renilla luciferase control plasmid (pRL-TK). Forty-eight hours after transfection, the cells were lysed and assessed for luciferase activity. Data are presented as the ratio of firefly luciferase to renilla. Different letters above the bars indicate statistically different means by ANOVA and post hoc analysis (n=4; P< 0.05).

Journal: Oncogene

Article Title: BMP- specific SMADs function as novel repressors of PDGFA and modulate its expression in ovarian granulosa cells and tumors

doi: 10.1038/onc.2012.392

Figure Lengend Snippet: SMAD1/5 expression antagonizes Sp1 induction of PDGFA promoter. (A) COV434 cells were transiently co-transfected with 1 μg of -881:luc of the human PDGFA promoter and 0.5 μg of Sp1 and SMAD1/5 along with the renilla luciferase control plasmid (pRL-TK). Forty-eight hours after transfection, the cells were lysed and assessed for luciferase activity. Data are presented as the ratio of firefly luciferase to renilla. (B) Representative Western blot of whole cell lysates from COV434 cells transfected with either pcDNA3.1 or Flag-tagged SMAD1/5 expression plasmids and blotted with mouse anti-FLAG M2 antibody and mouse anti-β-actin antibody (loading control). (C) ChIP analysis in COV434 cells transfected with pcDNA3.1 or Flag-tagged SMAD1/5 expression plasmids. Chromatin cross-linked protein DNA complexes were immunoprecipitated with either anti-Sp1 antibody or with non-specific IgG and the PDGF-A promoter amplified by real-time PCR using locus specific primers. (D) SMAD1/5 represses the wild-type PDGFA promoter (-261:luc) but not the promoter bearing a mutation in the SMAD1/5 binding site (-261mutA: luc). COV434 cells were transiently co-transfected with 1 μg of -261:luc (control) or -261mutA: luc (mutant) and 0.5 μg of Sp1 and SMAD1/5 along with the renilla luciferase control plasmid (pRL-TK). Forty-eight hours after transfection, the cells were lysed and assessed for luciferase activity. Data are presented as the ratio of firefly luciferase to renilla. Different letters above the bars indicate statistically different means by ANOVA and post hoc analysis (n=4; P< 0.05).

Techniques: Expressing, Transfection, Luciferase, Control, Plasmid Preparation, Activity Assay, Western Blot, Immunoprecipitation, Amplification, Real-time Polymerase Chain Reaction, Mutagenesis, Binding Assay

Model of BR-Smad repression of PDGFA promoter activity. Earlier studies demonstrated that Sp1 mediates the basal transcription of the PDGFA gene through binding to four consensus binding sites in the proximal promoter region (denoted sites “A–D”). Based on ChIP data, we suggest that SMAD1 and SMAD5 bind to the proximal promoter region of PDGFA and block the “A” site for the Sp1 binding, thereby inhibiting gene expression. Loss of the BR-SMADs ( SMAD1 and SMAD5 ) in ovarian granulosa cells thus allows Sp1 to bind to the “A” site and leads to increased PDGFA activation, possibly initiated cell proliferation and contributing to disease onset.

Journal: Oncogene

Article Title: BMP- specific SMADs function as novel repressors of PDGFA and modulate its expression in ovarian granulosa cells and tumors

doi: 10.1038/onc.2012.392

Figure Lengend Snippet: Model of BR-Smad repression of PDGFA promoter activity. Earlier studies demonstrated that Sp1 mediates the basal transcription of the PDGFA gene through binding to four consensus binding sites in the proximal promoter region (denoted sites “A–D”). Based on ChIP data, we suggest that SMAD1 and SMAD5 bind to the proximal promoter region of PDGFA and block the “A” site for the Sp1 binding, thereby inhibiting gene expression. Loss of the BR-SMADs ( SMAD1 and SMAD5 ) in ovarian granulosa cells thus allows Sp1 to bind to the “A” site and leads to increased PDGFA activation, possibly initiated cell proliferation and contributing to disease onset.

Techniques: Activity Assay, Binding Assay, Blocking Assay, Gene Expression, Activation Assay

Review

ebm basal medium (Chem Impex International)

Structured Review

Images

Related Articles

Cell Culture:

Similar Products

Image Search Results