Structured Review

Santa Cruz Biotechnology cygb
Ngb is an axon-regeneration inducer in primary cultured neurons after OGD/Re. a Western blotting analysis of Ngb and GAP43 in primary cultures of mouse cerebral cortical neurons. Cultured neurons at DIV 7 were incubated with OGD media at 1% O 2 for 1 h and then re-incubated with normal neurobasal media at 21% O 2 for various times (OGD/Re). Total soluble proteins were extracted and subjected to western blotting analysis with <t>anti-Ngb/Cygb/GAP43/β-tubulin</t> antibodies. Statistical analysis demonstrated that Ngb and GAP43 were significantly upregulated at 6, 12, and 24 h of OGD/Re. Data are presented as means ± S.E.M. * P
Cygb, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Neuroglobin boosts axon regeneration during ischemic reperfusion via p38 binding and activation depending on oxygen signal"

Article Title: Neuroglobin boosts axon regeneration during ischemic reperfusion via p38 binding and activation depending on oxygen signal

Journal: Cell Death & Disease

doi: 10.1038/s41419-017-0260-8

Ngb is an axon-regeneration inducer in primary cultured neurons after OGD/Re. a Western blotting analysis of Ngb and GAP43 in primary cultures of mouse cerebral cortical neurons. Cultured neurons at DIV 7 were incubated with OGD media at 1% O 2 for 1 h and then re-incubated with normal neurobasal media at 21% O 2 for various times (OGD/Re). Total soluble proteins were extracted and subjected to western blotting analysis with anti-Ngb/Cygb/GAP43/β-tubulin antibodies. Statistical analysis demonstrated that Ngb and GAP43 were significantly upregulated at 6, 12, and 24 h of OGD/Re. Data are presented as means ± S.E.M. * P
Figure Legend Snippet: Ngb is an axon-regeneration inducer in primary cultured neurons after OGD/Re. a Western blotting analysis of Ngb and GAP43 in primary cultures of mouse cerebral cortical neurons. Cultured neurons at DIV 7 were incubated with OGD media at 1% O 2 for 1 h and then re-incubated with normal neurobasal media at 21% O 2 for various times (OGD/Re). Total soluble proteins were extracted and subjected to western blotting analysis with anti-Ngb/Cygb/GAP43/β-tubulin antibodies. Statistical analysis demonstrated that Ngb and GAP43 were significantly upregulated at 6, 12, and 24 h of OGD/Re. Data are presented as means ± S.E.M. * P

Techniques Used: Cell Culture, Western Blot, Incubation

2) Product Images from "Cytoglobin: a potential marker for adipogenic differentiation in preadipocytes in vitro"

Article Title: Cytoglobin: a potential marker for adipogenic differentiation in preadipocytes in vitro

Journal: Cytotechnology

doi: 10.1007/s10616-016-0047-2

Western blot analysis of Cygb, PPARγ, CEBPα and FABP4. during differentiation process. a Western blot results of Cygb, PPARγ, CEBPα and FABP4 during differentiation process, b quantitative results of western blot band intensities. * P
Figure Legend Snippet: Western blot analysis of Cygb, PPARγ, CEBPα and FABP4. during differentiation process. a Western blot results of Cygb, PPARγ, CEBPα and FABP4 during differentiation process, b quantitative results of western blot band intensities. * P

Techniques Used: Western Blot

Cygb overexpression increases adipogenic differentiation. a Structure of Cygb gene construct, b relative mRNA expression level of Cygb in transduced cells, c oil red staining of differentiated 3T3-F442A–GFP and 3T3-F442A–Cygb cells, d gene expression levels of PPARγ, CEBPα and FABP4 in differentiated 3T3-F442A–GFP and 3T3-F442A–Cygb cells at the end of 8 day differentiation period. * P
Figure Legend Snippet: Cygb overexpression increases adipogenic differentiation. a Structure of Cygb gene construct, b relative mRNA expression level of Cygb in transduced cells, c oil red staining of differentiated 3T3-F442A–GFP and 3T3-F442A–Cygb cells, d gene expression levels of PPARγ, CEBPα and FABP4 in differentiated 3T3-F442A–GFP and 3T3-F442A–Cygb cells at the end of 8 day differentiation period. * P

Techniques Used: Over Expression, Construct, Expressing, Staining

Gene expression analysis of Cygb, PPARγ, CEBPα and FABP4 during differentiation process. Indicated genes showed an increased in relative mRNA expression in a time dependent manner. * P
Figure Legend Snippet: Gene expression analysis of Cygb, PPARγ, CEBPα and FABP4 during differentiation process. Indicated genes showed an increased in relative mRNA expression in a time dependent manner. * P

Techniques Used: Expressing

Immunocytochemistry analysis of PPARγ, CEBPα and FABP4 during differentiation process. PPARγ, CEBPα and FABP4 staining at day 0 and day 8 is shown. Expression of PPARγ, CEBPα and FABP4 increased during differentiation process. DAPI: Nuclei staining, FITC: Cygb staining. Scale bar 10 µm
Figure Legend Snippet: Immunocytochemistry analysis of PPARγ, CEBPα and FABP4 during differentiation process. PPARγ, CEBPα and FABP4 staining at day 0 and day 8 is shown. Expression of PPARγ, CEBPα and FABP4 increased during differentiation process. DAPI: Nuclei staining, FITC: Cygb staining. Scale bar 10 µm

Techniques Used: Immunocytochemistry, Staining, Expressing

3) Product Images from "Neuroglobin boosts axon regeneration during ischemic reperfusion via p38 binding and activation depending on oxygen signal"

Article Title: Neuroglobin boosts axon regeneration during ischemic reperfusion via p38 binding and activation depending on oxygen signal

Journal: Cell Death & Disease

doi: 10.1038/s41419-017-0260-8

Ngb is an axon-regeneration inducer in primary cultured neurons after OGD/Re. a Western blotting analysis of Ngb and GAP43 in primary cultures of mouse cerebral cortical neurons. Cultured neurons at DIV 7 were incubated with OGD media at 1% O 2 for 1 h and then re-incubated with normal neurobasal media at 21% O 2 for various times (OGD/Re). Total soluble proteins were extracted and subjected to western blotting analysis with anti-Ngb/Cygb/GAP43/β-tubulin antibodies. Statistical analysis demonstrated that Ngb and GAP43 were significantly upregulated at 6, 12, and 24 h of OGD/Re. Data are presented as means ± S.E.M. * P
Figure Legend Snippet: Ngb is an axon-regeneration inducer in primary cultured neurons after OGD/Re. a Western blotting analysis of Ngb and GAP43 in primary cultures of mouse cerebral cortical neurons. Cultured neurons at DIV 7 were incubated with OGD media at 1% O 2 for 1 h and then re-incubated with normal neurobasal media at 21% O 2 for various times (OGD/Re). Total soluble proteins were extracted and subjected to western blotting analysis with anti-Ngb/Cygb/GAP43/β-tubulin antibodies. Statistical analysis demonstrated that Ngb and GAP43 were significantly upregulated at 6, 12, and 24 h of OGD/Re. Data are presented as means ± S.E.M. * P

Techniques Used: Cell Culture, Western Blot, Incubation

4) Product Images from "The epigenetically downregulated factor CYGB suppresses breast cancer through inhibition of glucose metabolism"

Article Title: The epigenetically downregulated factor CYGB suppresses breast cancer through inhibition of glucose metabolism

Journal: Journal of Experimental & Clinical Cancer Research : CR

doi: 10.1186/s13046-018-0979-9

Expression and methylation of CYGB in breast cancer. a Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis of CYGB mRNA expression in breast tumor tissue samples and paired tumor adjacent tissue samples. b Low CYGB expression is correlated with poor ten-year relapse-free survival (RFS) in breast cancer patients. Prognosis data was acquired and analyzed using the Kaplan-Meier Plotter database ( www.kmplot.com/analysis/index. php?p=service cancer=breast ). c CYGB mRNA expression and promoter methylation in paired breast cancer tissue samples and non-cancerous breast tissue samples of 36 patients from The Cancer Genome Atlas (TCGA). The data were accessed through cBioPortal ( http://www.cbioportal.org /). d RT-PCR and MSP analyses of CYGB mRNA expression and promoter methylation in breast cancer cell lines. Non-tumorigenic mammary epithelial cell lines MCF10A and HMEC as well as normal breast tissue samples were used as controls. GAPDH was detected as an input control. e RT-PCR and MSP indicate demethylation by Aza and TSA (A + T) restored CYGB expression in BT549, MB231, MCF7 cells. GAPDH was detected as an input control. Aza: 5-aza-2′-deoxycytidine; BN: breast normal tissue; BrCa: breast cancer tissue; M: methylated; MSP: methylation-specific polymerase chain reaction; RT-PCR: reverse transcription-polymerase chain reaction; U: unmethylated
Figure Legend Snippet: Expression and methylation of CYGB in breast cancer. a Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis of CYGB mRNA expression in breast tumor tissue samples and paired tumor adjacent tissue samples. b Low CYGB expression is correlated with poor ten-year relapse-free survival (RFS) in breast cancer patients. Prognosis data was acquired and analyzed using the Kaplan-Meier Plotter database ( www.kmplot.com/analysis/index. php?p=service cancer=breast ). c CYGB mRNA expression and promoter methylation in paired breast cancer tissue samples and non-cancerous breast tissue samples of 36 patients from The Cancer Genome Atlas (TCGA). The data were accessed through cBioPortal ( http://www.cbioportal.org /). d RT-PCR and MSP analyses of CYGB mRNA expression and promoter methylation in breast cancer cell lines. Non-tumorigenic mammary epithelial cell lines MCF10A and HMEC as well as normal breast tissue samples were used as controls. GAPDH was detected as an input control. e RT-PCR and MSP indicate demethylation by Aza and TSA (A + T) restored CYGB expression in BT549, MB231, MCF7 cells. GAPDH was detected as an input control. Aza: 5-aza-2′-deoxycytidine; BN: breast normal tissue; BrCa: breast cancer tissue; M: methylated; MSP: methylation-specific polymerase chain reaction; RT-PCR: reverse transcription-polymerase chain reaction; U: unmethylated

Techniques Used: Expressing, Methylation, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Polymerase Chain Reaction

Related Articles

Electrophoresis:

Article Title: Cytoglobin expression in the hepatic stellate cell line HSC-T6 is regulated by extracellular matrix proteins dependent on FAK-signalling
Article Snippet: .. 20 μg) determined by the method of Bradford [ ], were prepared in Laemmli loading buffer (0.42 M SDS, 0.87 mM bromophenol blue, 47 % v /v glycerol, 60 mM Tris pH 6.8 and 1.6 M β-mercaptoethanol) and resolved on a 12.5 % SDS-PAGE electrophoresis gel, transferred to PVDF and probed for Cygb (Santa-Cruz, clone FL-190, 1:200 dilution). .. Equal loading was confirmed by blotting with β-actin (Sigma, 1:10,000 dilution), secondary antibody (goat anti-rabbit or goat anti-mouse HRP, DAKO) was used at 1:500 or 1:1000 and the signal was visualised using ECL femto reagent (Geneflow) and an X-ograph (AGFA Curix60).

Western Blot:

Article Title: Cytoglobin: a potential marker for adipogenic differentiation in preadipocytes in vitro
Article Snippet: .. Primary antibodies against PPARγ (1:1000 dilution; MA5-14889; Sigma), CEBPα (1:1000 dilution; 8178; Cell Signaling Technology), FABP4 (1:1000 dilution; sc-271529; Santa Cruz Biotechnology), and Cygb (1:1000 dilution; sc-271529; Santa Cruz Biotechnology) were used for western blot analysis. .. Total protein was isolated from cells at day 0, 2, 4 and 8 of differentiation using RIPA Buffer (sc-24948, Santa Cruz Biotechnology) and protein concentrations were estimated using BCA assay (23227, Pierce, Rockford, IL, USA).

Incubation:

Article Title: Mechanisms of Neuroprotection from Hypoxia-Ischemia (HI) Brain Injury by Up-regulation of Cytoglobin (CYGB) in a Neonatal Rat Model *
Article Snippet: .. The membrane was blocked by a 1-h incubation at room temperature in a Tris-buffered saline solution (TBS-T; 20 m m Tris, pH 7.6, 135 m m NaCl, and 0.05% Tween) containing 5% nonfat dry milk and then incubated with different primary antibodies, including anti-CYGB (1:400 dilution; Santa Cruz Biotechnology, Inc.), anti-β-actin (1:1000 dilution; CST) overnight at 4 °C. .. After washing the membrane three times with TBS-T, the secondary antibody HRP-labeled goat anti-rabbit IgG was then added to the membrane according to the vendor's recommendation (1:8000 dilution; CST) and incubated for 1 h at room temperature and then washed again as described previously.

Article Title: Cytoglobin: a potential marker for adipogenic differentiation in preadipocytes in vitro
Article Snippet: .. Cells were incubated with the following primary antibodies; anti-PPARγ (1:200 dilution; MA5-14889; Sigma), anti-CEBPα (1:400 dilution; 8178; Cell Signaling Technology, Beverly, MA, USA), anti-FABP4 (1:100 dilution; sc-271529; Santa Cruz Biotechnology), and anti-Cygb (1:200 dilution; sc-66855; Santa Cruz Biotechnology) overnight at 4 °C. .. Cells were then treated with AlexaFluor 488 FITC conjugated goat anti-rabbit or anti-mouse IgG (1:200 dilution; ThermoFisher, Waltham, MA, USA) secondary antibodies for 1 h at room temperature.

SDS Page:

Article Title: Cytoglobin expression in the hepatic stellate cell line HSC-T6 is regulated by extracellular matrix proteins dependent on FAK-signalling
Article Snippet: .. 20 μg) determined by the method of Bradford [ ], were prepared in Laemmli loading buffer (0.42 M SDS, 0.87 mM bromophenol blue, 47 % v /v glycerol, 60 mM Tris pH 6.8 and 1.6 M β-mercaptoethanol) and resolved on a 12.5 % SDS-PAGE electrophoresis gel, transferred to PVDF and probed for Cygb (Santa-Cruz, clone FL-190, 1:200 dilution). .. Equal loading was confirmed by blotting with β-actin (Sigma, 1:10,000 dilution), secondary antibody (goat anti-rabbit or goat anti-mouse HRP, DAKO) was used at 1:500 or 1:1000 and the signal was visualised using ECL femto reagent (Geneflow) and an X-ograph (AGFA Curix60).

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    Santa Cruz Biotechnology cygb
    Ngb is an axon-regeneration inducer in primary cultured neurons after OGD/Re. a Western blotting analysis of Ngb and GAP43 in primary cultures of mouse cerebral cortical neurons. Cultured neurons at DIV 7 were incubated with OGD media at 1% O 2 for 1 h and then re-incubated with normal neurobasal media at 21% O 2 for various times (OGD/Re). Total soluble proteins were extracted and subjected to western blotting analysis with <t>anti-Ngb/Cygb/GAP43/β-tubulin</t> antibodies. Statistical analysis demonstrated that Ngb and GAP43 were significantly upregulated at 6, 12, and 24 h of OGD/Re. Data are presented as means ± S.E.M. * P
    Cygb, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cygb/product/Santa Cruz Biotechnology
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cygb - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    90
    Santa Cruz Biotechnology anti cygb
    The expression of <t>Cygb</t> in HI animals injected with shRNA or cDNA. The expression of Cygb mRNA at 18 h post-HI was confirmed by RT-PCR ( A ). The expression of CYGB protein at 24 h post-HI was confirmed by Western blotting using anti-CYGB antibody ( C ). <t>β-Actin</t>
    Anti Cygb, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cygb/product/Santa Cruz Biotechnology
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    anti cygb - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

    Image Search Results


    Ngb is an axon-regeneration inducer in primary cultured neurons after OGD/Re. a Western blotting analysis of Ngb and GAP43 in primary cultures of mouse cerebral cortical neurons. Cultured neurons at DIV 7 were incubated with OGD media at 1% O 2 for 1 h and then re-incubated with normal neurobasal media at 21% O 2 for various times (OGD/Re). Total soluble proteins were extracted and subjected to western blotting analysis with anti-Ngb/Cygb/GAP43/β-tubulin antibodies. Statistical analysis demonstrated that Ngb and GAP43 were significantly upregulated at 6, 12, and 24 h of OGD/Re. Data are presented as means ± S.E.M. * P

    Journal: Cell Death & Disease

    Article Title: Neuroglobin boosts axon regeneration during ischemic reperfusion via p38 binding and activation depending on oxygen signal

    doi: 10.1038/s41419-017-0260-8

    Figure Lengend Snippet: Ngb is an axon-regeneration inducer in primary cultured neurons after OGD/Re. a Western blotting analysis of Ngb and GAP43 in primary cultures of mouse cerebral cortical neurons. Cultured neurons at DIV 7 were incubated with OGD media at 1% O 2 for 1 h and then re-incubated with normal neurobasal media at 21% O 2 for various times (OGD/Re). Total soluble proteins were extracted and subjected to western blotting analysis with anti-Ngb/Cygb/GAP43/β-tubulin antibodies. Statistical analysis demonstrated that Ngb and GAP43 were significantly upregulated at 6, 12, and 24 h of OGD/Re. Data are presented as means ± S.E.M. * P

    Article Snippet: Antibodies against Ngb, Flag (Sigma, Saint Louis, MO, USA), GAP43, GFAP, NeuN, NF200, p-p38, p38 (Cell Signaling Technology, Boston, USA), β-tubulin, green fluorescent protein (GFP), Cygb, GST, His (Santa Cruz Biotechnology, Santa Cruz, USA), and Tau-1 (Merk Millipore Ltd., Darmstadt, Germany) were purchased.

    Techniques: Cell Culture, Western Blot, Incubation

    Western blot analysis of Cygb, PPARγ, CEBPα and FABP4. during differentiation process. a Western blot results of Cygb, PPARγ, CEBPα and FABP4 during differentiation process, b quantitative results of western blot band intensities. * P

    Journal: Cytotechnology

    Article Title: Cytoglobin: a potential marker for adipogenic differentiation in preadipocytes in vitro

    doi: 10.1007/s10616-016-0047-2

    Figure Lengend Snippet: Western blot analysis of Cygb, PPARγ, CEBPα and FABP4. during differentiation process. a Western blot results of Cygb, PPARγ, CEBPα and FABP4 during differentiation process, b quantitative results of western blot band intensities. * P

    Article Snippet: Primary antibodies against PPARγ (1:1000 dilution; MA5-14889; Sigma), CEBPα (1:1000 dilution; 8178; Cell Signaling Technology), FABP4 (1:1000 dilution; sc-271529; Santa Cruz Biotechnology), and Cygb (1:1000 dilution; sc-271529; Santa Cruz Biotechnology) were used for western blot analysis.

    Techniques: Western Blot

    Cygb overexpression increases adipogenic differentiation. a Structure of Cygb gene construct, b relative mRNA expression level of Cygb in transduced cells, c oil red staining of differentiated 3T3-F442A–GFP and 3T3-F442A–Cygb cells, d gene expression levels of PPARγ, CEBPα and FABP4 in differentiated 3T3-F442A–GFP and 3T3-F442A–Cygb cells at the end of 8 day differentiation period. * P

    Journal: Cytotechnology

    Article Title: Cytoglobin: a potential marker for adipogenic differentiation in preadipocytes in vitro

    doi: 10.1007/s10616-016-0047-2

    Figure Lengend Snippet: Cygb overexpression increases adipogenic differentiation. a Structure of Cygb gene construct, b relative mRNA expression level of Cygb in transduced cells, c oil red staining of differentiated 3T3-F442A–GFP and 3T3-F442A–Cygb cells, d gene expression levels of PPARγ, CEBPα and FABP4 in differentiated 3T3-F442A–GFP and 3T3-F442A–Cygb cells at the end of 8 day differentiation period. * P

    Article Snippet: Primary antibodies against PPARγ (1:1000 dilution; MA5-14889; Sigma), CEBPα (1:1000 dilution; 8178; Cell Signaling Technology), FABP4 (1:1000 dilution; sc-271529; Santa Cruz Biotechnology), and Cygb (1:1000 dilution; sc-271529; Santa Cruz Biotechnology) were used for western blot analysis.

    Techniques: Over Expression, Construct, Expressing, Staining

    Gene expression analysis of Cygb, PPARγ, CEBPα and FABP4 during differentiation process. Indicated genes showed an increased in relative mRNA expression in a time dependent manner. * P

    Journal: Cytotechnology

    Article Title: Cytoglobin: a potential marker for adipogenic differentiation in preadipocytes in vitro

    doi: 10.1007/s10616-016-0047-2

    Figure Lengend Snippet: Gene expression analysis of Cygb, PPARγ, CEBPα and FABP4 during differentiation process. Indicated genes showed an increased in relative mRNA expression in a time dependent manner. * P

    Article Snippet: Primary antibodies against PPARγ (1:1000 dilution; MA5-14889; Sigma), CEBPα (1:1000 dilution; 8178; Cell Signaling Technology), FABP4 (1:1000 dilution; sc-271529; Santa Cruz Biotechnology), and Cygb (1:1000 dilution; sc-271529; Santa Cruz Biotechnology) were used for western blot analysis.

    Techniques: Expressing

    Immunocytochemistry analysis of PPARγ, CEBPα and FABP4 during differentiation process. PPARγ, CEBPα and FABP4 staining at day 0 and day 8 is shown. Expression of PPARγ, CEBPα and FABP4 increased during differentiation process. DAPI: Nuclei staining, FITC: Cygb staining. Scale bar 10 µm

    Journal: Cytotechnology

    Article Title: Cytoglobin: a potential marker for adipogenic differentiation in preadipocytes in vitro

    doi: 10.1007/s10616-016-0047-2

    Figure Lengend Snippet: Immunocytochemistry analysis of PPARγ, CEBPα and FABP4 during differentiation process. PPARγ, CEBPα and FABP4 staining at day 0 and day 8 is shown. Expression of PPARγ, CEBPα and FABP4 increased during differentiation process. DAPI: Nuclei staining, FITC: Cygb staining. Scale bar 10 µm

    Article Snippet: Primary antibodies against PPARγ (1:1000 dilution; MA5-14889; Sigma), CEBPα (1:1000 dilution; 8178; Cell Signaling Technology), FABP4 (1:1000 dilution; sc-271529; Santa Cruz Biotechnology), and Cygb (1:1000 dilution; sc-271529; Santa Cruz Biotechnology) were used for western blot analysis.

    Techniques: Immunocytochemistry, Staining, Expressing

    Ngb is an axon-regeneration inducer in primary cultured neurons after OGD/Re. a Western blotting analysis of Ngb and GAP43 in primary cultures of mouse cerebral cortical neurons. Cultured neurons at DIV 7 were incubated with OGD media at 1% O 2 for 1 h and then re-incubated with normal neurobasal media at 21% O 2 for various times (OGD/Re). Total soluble proteins were extracted and subjected to western blotting analysis with anti-Ngb/Cygb/GAP43/β-tubulin antibodies. Statistical analysis demonstrated that Ngb and GAP43 were significantly upregulated at 6, 12, and 24 h of OGD/Re. Data are presented as means ± S.E.M. * P

    Journal: Cell Death & Disease

    Article Title: Neuroglobin boosts axon regeneration during ischemic reperfusion via p38 binding and activation depending on oxygen signal

    doi: 10.1038/s41419-017-0260-8

    Figure Lengend Snippet: Ngb is an axon-regeneration inducer in primary cultured neurons after OGD/Re. a Western blotting analysis of Ngb and GAP43 in primary cultures of mouse cerebral cortical neurons. Cultured neurons at DIV 7 were incubated with OGD media at 1% O 2 for 1 h and then re-incubated with normal neurobasal media at 21% O 2 for various times (OGD/Re). Total soluble proteins were extracted and subjected to western blotting analysis with anti-Ngb/Cygb/GAP43/β-tubulin antibodies. Statistical analysis demonstrated that Ngb and GAP43 were significantly upregulated at 6, 12, and 24 h of OGD/Re. Data are presented as means ± S.E.M. * P

    Article Snippet: Antibodies against Ngb, Flag (Sigma, Saint Louis, MO, USA), GAP43, GFAP, NeuN, NF200, p-p38, p38 (Cell Signaling Technology, Boston, USA), β-tubulin, green fluorescent protein (GFP), Cygb, GST, His (Santa Cruz Biotechnology, Santa Cruz, USA), and Tau-1 (Merk Millipore Ltd., Darmstadt, Germany) were purchased.

    Techniques: Cell Culture, Western Blot, Incubation

    The expression of Cygb in HI animals injected with shRNA or cDNA. The expression of Cygb mRNA at 18 h post-HI was confirmed by RT-PCR ( A ). The expression of CYGB protein at 24 h post-HI was confirmed by Western blotting using anti-CYGB antibody ( C ). β-Actin

    Journal: The Journal of Biological Chemistry

    Article Title: Mechanisms of Neuroprotection from Hypoxia-Ischemia (HI) Brain Injury by Up-regulation of Cytoglobin (CYGB) in a Neonatal Rat Model *

    doi: 10.1074/jbc.M112.428789

    Figure Lengend Snippet: The expression of Cygb in HI animals injected with shRNA or cDNA. The expression of Cygb mRNA at 18 h post-HI was confirmed by RT-PCR ( A ). The expression of CYGB protein at 24 h post-HI was confirmed by Western blotting using anti-CYGB antibody ( C ). β-Actin

    Article Snippet: The membrane was blocked by a 1-h incubation at room temperature in a Tris-buffered saline solution (TBS-T; 20 m m Tris, pH 7.6, 135 m m NaCl, and 0.05% Tween) containing 5% nonfat dry milk and then incubated with different primary antibodies, including anti-CYGB (1:400 dilution; Santa Cruz Biotechnology, Inc.), anti-β-actin (1:1000 dilution; CST) overnight at 4 °C.

    Techniques: Expressing, Injection, shRNA, Reverse Transcription Polymerase Chain Reaction, Western Blot

    Infection of neonatal brain with adenovirus carrying Cygb cDNA or shRNA. CYGB expression at various time points was confirmed by Western blotting using anti-CYGB antibody. β-actin was used as an internal control. A and C , representative electrophoretic

    Journal: The Journal of Biological Chemistry

    Article Title: Mechanisms of Neuroprotection from Hypoxia-Ischemia (HI) Brain Injury by Up-regulation of Cytoglobin (CYGB) in a Neonatal Rat Model *

    doi: 10.1074/jbc.M112.428789

    Figure Lengend Snippet: Infection of neonatal brain with adenovirus carrying Cygb cDNA or shRNA. CYGB expression at various time points was confirmed by Western blotting using anti-CYGB antibody. β-actin was used as an internal control. A and C , representative electrophoretic

    Article Snippet: The membrane was blocked by a 1-h incubation at room temperature in a Tris-buffered saline solution (TBS-T; 20 m m Tris, pH 7.6, 135 m m NaCl, and 0.05% Tween) containing 5% nonfat dry milk and then incubated with different primary antibodies, including anti-CYGB (1:400 dilution; Santa Cruz Biotechnology, Inc.), anti-β-actin (1:1000 dilution; CST) overnight at 4 °C.

    Techniques: Infection, shRNA, Expressing, Western Blot