probdnf  (Alomone Labs)


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    Alomone Labs probdnf
    Probdnf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
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    probdnf - by Bioz Stars, 2023-01
    94/100 stars

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    probdnf  (Alomone Labs)


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  • 94

    Structured Review

    Alomone Labs probdnf
    Probdnf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/probdnf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    probdnf - by Bioz Stars, 2023-01
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    human pro bdnf biotin  (Alomone Labs)


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  • 94

    Structured Review

    Alomone Labs human pro bdnf biotin
    Human Pro Bdnf Biotin, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human pro bdnf biotin/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
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    94/100 stars

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    human dlg2 protein  (Alomone Labs)


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  • 92

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    Alomone Labs human dlg2 protein
    LncRNA <t>DLG2-AS1</t> is downregulated in lung adenocarcinoma (LUAD). ( a ) heatmap of the 90 lncRNAs analyzed with the Human LncRNA Profile Kit from System Biosciences (SBI); ( b ) DLG2-AS1 expression in normal and tumor samples; and ( c ) tumor/normal fold change (FC(T/N)) of DLG2-AS1 expression in the 65 LUAD patients. Shown in a darker color are the patients who presented DLG2-AS1 downregulation (FC(T/N) < 0.66). *** = p -value < 0.001.
    Human Dlg2 Protein, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 1 article reviews
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    human dlg2 protein - by Bioz Stars, 2023-01
    92/100 stars

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    1) Product Images from "LncRNA DLG2-AS1 as a Novel Biomarker in Lung Adenocarcinoma"

    Article Title: LncRNA DLG2-AS1 as a Novel Biomarker in Lung Adenocarcinoma

    Journal: Cancers

    doi: 10.3390/cancers12082080

    LncRNA DLG2-AS1 is downregulated in lung adenocarcinoma (LUAD). ( a ) heatmap of the 90 lncRNAs analyzed with the Human LncRNA Profile Kit from System Biosciences (SBI); ( b ) DLG2-AS1 expression in normal and tumor samples; and ( c ) tumor/normal fold change (FC(T/N)) of DLG2-AS1 expression in the 65 LUAD patients. Shown in a darker color are the patients who presented DLG2-AS1 downregulation (FC(T/N) < 0.66). *** = p -value < 0.001.
    Figure Legend Snippet: LncRNA DLG2-AS1 is downregulated in lung adenocarcinoma (LUAD). ( a ) heatmap of the 90 lncRNAs analyzed with the Human LncRNA Profile Kit from System Biosciences (SBI); ( b ) DLG2-AS1 expression in normal and tumor samples; and ( c ) tumor/normal fold change (FC(T/N)) of DLG2-AS1 expression in the 65 LUAD patients. Shown in a darker color are the patients who presented DLG2-AS1 downregulation (FC(T/N) < 0.66). *** = p -value < 0.001.

    Techniques Used: Expressing

    DLG2-AS1 restoration model on lung adenocarcinoma (LUAD) cell lines successfully overexpresses DLG2-AS1. DLG2-AS1 relative expression was measured by RT-qPCR 5 days after transfection, and normalized by the expression of the reference gene U1 snRNA. In all three cell lines tested, DLG2-AS1 was successfully overexpressed (relative expression (pLVX-DLG2AS/EV) > 2). * = p -value < 0.05.
    Figure Legend Snippet: DLG2-AS1 restoration model on lung adenocarcinoma (LUAD) cell lines successfully overexpresses DLG2-AS1. DLG2-AS1 relative expression was measured by RT-qPCR 5 days after transfection, and normalized by the expression of the reference gene U1 snRNA. In all three cell lines tested, DLG2-AS1 was successfully overexpressed (relative expression (pLVX-DLG2AS/EV) > 2). * = p -value < 0.05.

    Techniques Used: Expressing, Quantitative RT-PCR, Transfection

    DLG2-AS1 is not a cis-regulator of DLG2 expression. ( a ) RNA expression levels of DLG2 were unaffected after DLG2-AS1 overexpression (pLVX-DLG2AS1) compared to empty vector transfection (EV). Results from RT-qPCR were normalized to expression of the reference gene U1 snRNA. ( b ) Protein levels of human DLG2 did not change upon transfection with pLVX-DLG2AS1 compared to EV in two independent Western blot experiments (relative pLVX-DLG2AS1/EV densitometry measure mean = 0.984 ± 0.199, p = 0.929). Uncropped WB images are available at the .
    Figure Legend Snippet: DLG2-AS1 is not a cis-regulator of DLG2 expression. ( a ) RNA expression levels of DLG2 were unaffected after DLG2-AS1 overexpression (pLVX-DLG2AS1) compared to empty vector transfection (EV). Results from RT-qPCR were normalized to expression of the reference gene U1 snRNA. ( b ) Protein levels of human DLG2 did not change upon transfection with pLVX-DLG2AS1 compared to EV in two independent Western blot experiments (relative pLVX-DLG2AS1/EV densitometry measure mean = 0.984 ± 0.199, p = 0.929). Uncropped WB images are available at the .

    Techniques Used: Expressing, RNA Expression, Over Expression, Plasmid Preparation, Transfection, Quantitative RT-PCR, Western Blot

    DLG2-AS1 shows potential as a lung adenocarcinoma (LUAD) biomarker: ( a ) receiver operating characteristic (ROC) curve and area under curve (AUC) value of DLG2-AS1 expression in our cohort of LUAD patients; and ( b ) ROC curves and AUC values of other LUAD biomarkers (EGFR, TP53, MALAT-1, and NEAT1), obtained from TCGA data.
    Figure Legend Snippet: DLG2-AS1 shows potential as a lung adenocarcinoma (LUAD) biomarker: ( a ) receiver operating characteristic (ROC) curve and area under curve (AUC) value of DLG2-AS1 expression in our cohort of LUAD patients; and ( b ) ROC curves and AUC values of other LUAD biomarkers (EGFR, TP53, MALAT-1, and NEAT1), obtained from TCGA data.

    Techniques Used: Biomarker Assay, Expressing

    Sequences of used oligonucleotides for qPCR and vector cloning.
    Figure Legend Snippet: Sequences of used oligonucleotides for qPCR and vector cloning.

    Techniques Used: Plasmid Preparation, Clone Assay, Sequencing

    nonglycosylated human probdnf  (Alomone Labs)


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    Alomone Labs nonglycosylated human probdnf
    <t>ProBDNF</t> stably expressed in HEK293F cells is glycosylated. N-Glycosylation of proBDNF and the prodomain is documented by mass shift following deglycosylation with PNGase F: A, detection of proBDNF and BDNF by an antibody recognizing the BDNF region; B, detection of proBDNF and the prodomain by an antibody recognizing the prodomain region. The gel images were spliced as indicated by space to exclude samples not related to the study.
    Nonglycosylated Human Probdnf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 1 article reviews
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    nonglycosylated human probdnf - by Bioz Stars, 2023-01
    92/100 stars

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    1) Product Images from "N -Glycosylation is required for secretion of the precursor to brain-derived neurotrophic factor (proBDNF) carrying sulfated LacdiNAc structures"

    Article Title: N -Glycosylation is required for secretion of the precursor to brain-derived neurotrophic factor (proBDNF) carrying sulfated LacdiNAc structures

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.RA119.009989

    ProBDNF stably expressed in HEK293F cells is glycosylated. N-Glycosylation of proBDNF and the prodomain is documented by mass shift following deglycosylation with PNGase F: A, detection of proBDNF and BDNF by an antibody recognizing the BDNF region; B, detection of proBDNF and the prodomain by an antibody recognizing the prodomain region. The gel images were spliced as indicated by space to exclude samples not related to the study.
    Figure Legend Snippet: ProBDNF stably expressed in HEK293F cells is glycosylated. N-Glycosylation of proBDNF and the prodomain is documented by mass shift following deglycosylation with PNGase F: A, detection of proBDNF and BDNF by an antibody recognizing the BDNF region; B, detection of proBDNF and the prodomain by an antibody recognizing the prodomain region. The gel images were spliced as indicated by space to exclude samples not related to the study.

    Techniques Used: Stable Transfection

    Extracted ion chromatograms (XIC) of (A) nonglycosylated, NYLDAANMSMR and (B) deglycosylated, NYLDAADMSMR glycopeptide of proBDNF secreted by HEK293F cells confirm site occupancy higher than 99%. C, tandem mass spectrum verifies deglycosylation of the proBDNF peptide under [18O]water.
    Figure Legend Snippet: Extracted ion chromatograms (XIC) of (A) nonglycosylated, NYLDAANMSMR and (B) deglycosylated, NYLDAADMSMR glycopeptide of proBDNF secreted by HEK293F cells confirm site occupancy higher than 99%. C, tandem mass spectrum verifies deglycosylation of the proBDNF peptide under [18O]water.

    Techniques Used:

    Expression of wildtype (WT) proBDNF and its N121Q mutant (NQ) in stably transfected HEK293F cells. Protein expression was determined by Western blotting using an antibody, recognizing both BDNF and proBDNF, in the conditioned media (A) and cell lysates (B). Arrows point to (pro)BDNF forms: glycosylated proBDNF (black arrow), nonglycosylated proBDNF (white arrow), and mature BDNF (gray arrow). M indicates MagicMark molecular weight marker. Expression of mRNA was determined in WT and N121Q mutant by RT-PCR (C). ProBDNF/BDNF expression in cell lysates (D) or conditioned media (E) of HEK293F cells expressing N121Q mutant treated with proteasome inhibitor MG132 (MG), lysosomal inhibitor chloroquine (CQ), or chemical chaperone 4-phenylbutyric acid (PBA). The black arrow points to the position of nonglycosylated proBDNF.
    Figure Legend Snippet: Expression of wildtype (WT) proBDNF and its N121Q mutant (NQ) in stably transfected HEK293F cells. Protein expression was determined by Western blotting using an antibody, recognizing both BDNF and proBDNF, in the conditioned media (A) and cell lysates (B). Arrows point to (pro)BDNF forms: glycosylated proBDNF (black arrow), nonglycosylated proBDNF (white arrow), and mature BDNF (gray arrow). M indicates MagicMark molecular weight marker. Expression of mRNA was determined in WT and N121Q mutant by RT-PCR (C). ProBDNF/BDNF expression in cell lysates (D) or conditioned media (E) of HEK293F cells expressing N121Q mutant treated with proteasome inhibitor MG132 (MG), lysosomal inhibitor chloroquine (CQ), or chemical chaperone 4-phenylbutyric acid (PBA). The black arrow points to the position of nonglycosylated proBDNF.

    Techniques Used: Expressing, Mutagenesis, Stable Transfection, Transfection, Western Blot, Molecular Weight, Marker, Reverse Transcription Polymerase Chain Reaction

    Kinetics of cleavage of glycosylated and nonglycosylated proBDNF by furin. Mass difference between glycosylated and nonglycosylated proBDNF enabled densitometric quantification of each proBDNF form: A, representative blot of the cleavage kinetic at the indicated time intervals; B, densitometric analysis of proBDNF intensities at the indicated time intervals. Results are expressed as percent of the proBDNF concentration (mean ± S.D., n = 3) at the beginning of the reaction. Note that proBDNF without C-terminal Myc-FLAG tag was used in the reaction as described under “Experimental procedures.”
    Figure Legend Snippet: Kinetics of cleavage of glycosylated and nonglycosylated proBDNF by furin. Mass difference between glycosylated and nonglycosylated proBDNF enabled densitometric quantification of each proBDNF form: A, representative blot of the cleavage kinetic at the indicated time intervals; B, densitometric analysis of proBDNF intensities at the indicated time intervals. Results are expressed as percent of the proBDNF concentration (mean ± S.D., n = 3) at the beginning of the reaction. Note that proBDNF without C-terminal Myc-FLAG tag was used in the reaction as described under “Experimental procedures.”

    Techniques Used: Concentration Assay, FLAG-tag

    Mass spectra of proBDNF under the following conditions. A, precursor profile of the tryptic glycopeptide NYLDAANM(ox)SM(ox)R of proBDNF expressed in HEK293F cells; B, HCD fragmentation spectrum of the major glycoform (m/z 1132.42) at low collision energy (10% NCE) showing that characteristic LacdiNAc oxonium ion HexNAc-HexNAc (m/z 407.17) and its fucosylated form (m/z 553.22) with complementary y-ions (m/z 1373, 1381, 1454, and 1555) dominate the composition.
    Figure Legend Snippet: Mass spectra of proBDNF under the following conditions. A, precursor profile of the tryptic glycopeptide NYLDAANM(ox)SM(ox)R of proBDNF expressed in HEK293F cells; B, HCD fragmentation spectrum of the major glycoform (m/z 1132.42) at low collision energy (10% NCE) showing that characteristic LacdiNAc oxonium ion HexNAc-HexNAc (m/z 407.17) and its fucosylated form (m/z 553.22) with complementary y-ions (m/z 1373, 1381, 1454, and 1555) dominate the composition.

    Techniques Used:

    Relative abundance of the glycoforms of HEK293F-produced proBDNF and prodomain expressed as % of all identified glycoforms
    Figure Legend Snippet: Relative abundance of the glycoforms of HEK293F-produced proBDNF and prodomain expressed as % of all identified glycoforms

    Techniques Used: Produced

    Extracted ion chromatograms of four of the most abundant glycopeptides of proBDNF expressed in PC12 cells. A, C, and D, LacdiNAc containing structures; B, LacNAc containing structure. The arrow points to the peaks of glycopeptides aligned with slight retention time shifts as expected based on the hydrophilicity of the attached glycan. Numbers represent intensity of the extracted peaks (×105).
    Figure Legend Snippet: Extracted ion chromatograms of four of the most abundant glycopeptides of proBDNF expressed in PC12 cells. A, C, and D, LacdiNAc containing structures; B, LacNAc containing structure. The arrow points to the peaks of glycopeptides aligned with slight retention time shifts as expected based on the hydrophilicity of the attached glycan. Numbers represent intensity of the extracted peaks (×105).

    Techniques Used:

    Impact of modulators of glycosylation on the BDNF/proBDNF ratio in HEK293F cells. Western blotting using antibodies to prodomain and mature BDNF regions was analyzed by densitometry for the following: A, intracellular BDNF to proBDNF ratio; B, secreted BDNF to proBDNF ratio; C, intracellular prodomain to proBDNF ratio; and D, secreted prodomain to proBDNF ratio. The representative Western blotting is labeled as the graphs: CTRL, control; CHLOR, 50 mm sodium chlorate; KIF, 1 μg/ml kifunensine; TNMC, 5 μg/ml of tunicamycin. Results are expressed as scatter plot (mean ± S.D., n = 3). *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared with CTRL, one-way analysis of variance with Bonferroni adjustment.
    Figure Legend Snippet: Impact of modulators of glycosylation on the BDNF/proBDNF ratio in HEK293F cells. Western blotting using antibodies to prodomain and mature BDNF regions was analyzed by densitometry for the following: A, intracellular BDNF to proBDNF ratio; B, secreted BDNF to proBDNF ratio; C, intracellular prodomain to proBDNF ratio; and D, secreted prodomain to proBDNF ratio. The representative Western blotting is labeled as the graphs: CTRL, control; CHLOR, 50 mm sodium chlorate; KIF, 1 μg/ml kifunensine; TNMC, 5 μg/ml of tunicamycin. Results are expressed as scatter plot (mean ± S.D., n = 3). *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared with CTRL, one-way analysis of variance with Bonferroni adjustment.

    Techniques Used: Western Blot, Labeling

    nonglycosylated human probdnf  (Alomone Labs)


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    Alomone Labs nonglycosylated human probdnf
    <t>ProBDNF</t> stably expressed in HEK293F cells is glycosylated. N-Glycosylation of proBDNF and the prodomain is documented by mass shift following deglycosylation with PNGase F: A, detection of proBDNF and BDNF by an antibody recognizing the BDNF region; B, detection of proBDNF and the prodomain by an antibody recognizing the prodomain region. The gel images were spliced as indicated by space to exclude samples not related to the study.
    Nonglycosylated Human Probdnf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nonglycosylated human probdnf/product/Alomone Labs
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nonglycosylated human probdnf - by Bioz Stars, 2023-01
    92/100 stars

    Images

    1) Product Images from "N -Glycosylation is required for secretion of the precursor to brain-derived neurotrophic factor (proBDNF) carrying sulfated LacdiNAc structures"

    Article Title: N -Glycosylation is required for secretion of the precursor to brain-derived neurotrophic factor (proBDNF) carrying sulfated LacdiNAc structures

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.RA119.009989

    ProBDNF stably expressed in HEK293F cells is glycosylated. N-Glycosylation of proBDNF and the prodomain is documented by mass shift following deglycosylation with PNGase F: A, detection of proBDNF and BDNF by an antibody recognizing the BDNF region; B, detection of proBDNF and the prodomain by an antibody recognizing the prodomain region. The gel images were spliced as indicated by space to exclude samples not related to the study.
    Figure Legend Snippet: ProBDNF stably expressed in HEK293F cells is glycosylated. N-Glycosylation of proBDNF and the prodomain is documented by mass shift following deglycosylation with PNGase F: A, detection of proBDNF and BDNF by an antibody recognizing the BDNF region; B, detection of proBDNF and the prodomain by an antibody recognizing the prodomain region. The gel images were spliced as indicated by space to exclude samples not related to the study.

    Techniques Used: Stable Transfection

    Extracted ion chromatograms (XIC) of (A) nonglycosylated, NYLDAANMSMR and (B) deglycosylated, NYLDAADMSMR glycopeptide of proBDNF secreted by HEK293F cells confirm site occupancy higher than 99%. C, tandem mass spectrum verifies deglycosylation of the proBDNF peptide under [18O]water.
    Figure Legend Snippet: Extracted ion chromatograms (XIC) of (A) nonglycosylated, NYLDAANMSMR and (B) deglycosylated, NYLDAADMSMR glycopeptide of proBDNF secreted by HEK293F cells confirm site occupancy higher than 99%. C, tandem mass spectrum verifies deglycosylation of the proBDNF peptide under [18O]water.

    Techniques Used:

    Expression of wildtype (WT) proBDNF and its N121Q mutant (NQ) in stably transfected HEK293F cells. Protein expression was determined by Western blotting using an antibody, recognizing both BDNF and proBDNF, in the conditioned media (A) and cell lysates (B). Arrows point to (pro)BDNF forms: glycosylated proBDNF (black arrow), nonglycosylated proBDNF (white arrow), and mature BDNF (gray arrow). M indicates MagicMark molecular weight marker. Expression of mRNA was determined in WT and N121Q mutant by RT-PCR (C). ProBDNF/BDNF expression in cell lysates (D) or conditioned media (E) of HEK293F cells expressing N121Q mutant treated with proteasome inhibitor MG132 (MG), lysosomal inhibitor chloroquine (CQ), or chemical chaperone 4-phenylbutyric acid (PBA). The black arrow points to the position of nonglycosylated proBDNF.
    Figure Legend Snippet: Expression of wildtype (WT) proBDNF and its N121Q mutant (NQ) in stably transfected HEK293F cells. Protein expression was determined by Western blotting using an antibody, recognizing both BDNF and proBDNF, in the conditioned media (A) and cell lysates (B). Arrows point to (pro)BDNF forms: glycosylated proBDNF (black arrow), nonglycosylated proBDNF (white arrow), and mature BDNF (gray arrow). M indicates MagicMark molecular weight marker. Expression of mRNA was determined in WT and N121Q mutant by RT-PCR (C). ProBDNF/BDNF expression in cell lysates (D) or conditioned media (E) of HEK293F cells expressing N121Q mutant treated with proteasome inhibitor MG132 (MG), lysosomal inhibitor chloroquine (CQ), or chemical chaperone 4-phenylbutyric acid (PBA). The black arrow points to the position of nonglycosylated proBDNF.

    Techniques Used: Expressing, Mutagenesis, Stable Transfection, Transfection, Western Blot, Molecular Weight, Marker, Reverse Transcription Polymerase Chain Reaction

    Kinetics of cleavage of glycosylated and nonglycosylated proBDNF by furin. Mass difference between glycosylated and nonglycosylated proBDNF enabled densitometric quantification of each proBDNF form: A, representative blot of the cleavage kinetic at the indicated time intervals; B, densitometric analysis of proBDNF intensities at the indicated time intervals. Results are expressed as percent of the proBDNF concentration (mean ± S.D., n = 3) at the beginning of the reaction. Note that proBDNF without C-terminal Myc-FLAG tag was used in the reaction as described under “Experimental procedures.”
    Figure Legend Snippet: Kinetics of cleavage of glycosylated and nonglycosylated proBDNF by furin. Mass difference between glycosylated and nonglycosylated proBDNF enabled densitometric quantification of each proBDNF form: A, representative blot of the cleavage kinetic at the indicated time intervals; B, densitometric analysis of proBDNF intensities at the indicated time intervals. Results are expressed as percent of the proBDNF concentration (mean ± S.D., n = 3) at the beginning of the reaction. Note that proBDNF without C-terminal Myc-FLAG tag was used in the reaction as described under “Experimental procedures.”

    Techniques Used: Concentration Assay, FLAG-tag

    Mass spectra of proBDNF under the following conditions. A, precursor profile of the tryptic glycopeptide NYLDAANM(ox)SM(ox)R of proBDNF expressed in HEK293F cells; B, HCD fragmentation spectrum of the major glycoform (m/z 1132.42) at low collision energy (10% NCE) showing that characteristic LacdiNAc oxonium ion HexNAc-HexNAc (m/z 407.17) and its fucosylated form (m/z 553.22) with complementary y-ions (m/z 1373, 1381, 1454, and 1555) dominate the composition.
    Figure Legend Snippet: Mass spectra of proBDNF under the following conditions. A, precursor profile of the tryptic glycopeptide NYLDAANM(ox)SM(ox)R of proBDNF expressed in HEK293F cells; B, HCD fragmentation spectrum of the major glycoform (m/z 1132.42) at low collision energy (10% NCE) showing that characteristic LacdiNAc oxonium ion HexNAc-HexNAc (m/z 407.17) and its fucosylated form (m/z 553.22) with complementary y-ions (m/z 1373, 1381, 1454, and 1555) dominate the composition.

    Techniques Used:

    Relative abundance of the glycoforms of HEK293F-produced proBDNF and prodomain expressed as % of all identified glycoforms
    Figure Legend Snippet: Relative abundance of the glycoforms of HEK293F-produced proBDNF and prodomain expressed as % of all identified glycoforms

    Techniques Used: Produced

    Extracted ion chromatograms of four of the most abundant glycopeptides of proBDNF expressed in PC12 cells. A, C, and D, LacdiNAc containing structures; B, LacNAc containing structure. The arrow points to the peaks of glycopeptides aligned with slight retention time shifts as expected based on the hydrophilicity of the attached glycan. Numbers represent intensity of the extracted peaks (×105).
    Figure Legend Snippet: Extracted ion chromatograms of four of the most abundant glycopeptides of proBDNF expressed in PC12 cells. A, C, and D, LacdiNAc containing structures; B, LacNAc containing structure. The arrow points to the peaks of glycopeptides aligned with slight retention time shifts as expected based on the hydrophilicity of the attached glycan. Numbers represent intensity of the extracted peaks (×105).

    Techniques Used:

    Impact of modulators of glycosylation on the BDNF/proBDNF ratio in HEK293F cells. Western blotting using antibodies to prodomain and mature BDNF regions was analyzed by densitometry for the following: A, intracellular BDNF to proBDNF ratio; B, secreted BDNF to proBDNF ratio; C, intracellular prodomain to proBDNF ratio; and D, secreted prodomain to proBDNF ratio. The representative Western blotting is labeled as the graphs: CTRL, control; CHLOR, 50 mm sodium chlorate; KIF, 1 μg/ml kifunensine; TNMC, 5 μg/ml of tunicamycin. Results are expressed as scatter plot (mean ± S.D., n = 3). *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared with CTRL, one-way analysis of variance with Bonferroni adjustment.
    Figure Legend Snippet: Impact of modulators of glycosylation on the BDNF/proBDNF ratio in HEK293F cells. Western blotting using antibodies to prodomain and mature BDNF regions was analyzed by densitometry for the following: A, intracellular BDNF to proBDNF ratio; B, secreted BDNF to proBDNF ratio; C, intracellular prodomain to proBDNF ratio; and D, secreted prodomain to proBDNF ratio. The representative Western blotting is labeled as the graphs: CTRL, control; CHLOR, 50 mm sodium chlorate; KIF, 1 μg/ml kifunensine; TNMC, 5 μg/ml of tunicamycin. Results are expressed as scatter plot (mean ± S.D., n = 3). *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared with CTRL, one-way analysis of variance with Bonferroni adjustment.

    Techniques Used: Western Blot, Labeling

    uncleavable probdnf  (Alomone Labs)


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    Alomone Labs uncleavable probdnf
    Uncleavable Probdnf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/uncleavable probdnf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    uncleavable probdnf - by Bioz Stars, 2023-01
    94/100 stars

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    mut probdnf  (Alomone Labs)


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    Alomone Labs mut probdnf
    Mut Probdnf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mut probdnf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
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    mut probdnf  (Alomone Labs)


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    Alomone Labs mut probdnf
    Mut Probdnf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    probdnf  (Alomone Labs)


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    Alomone Labs probdnf
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    Alomone Labs probdnf
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    Alomone Labs probdnf
    Probdnf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs human dlg2 protein
    LncRNA <t>DLG2-AS1</t> is downregulated in lung adenocarcinoma (LUAD). ( a ) heatmap of the 90 lncRNAs analyzed with the Human LncRNA Profile Kit from System Biosciences (SBI); ( b ) DLG2-AS1 expression in normal and tumor samples; and ( c ) tumor/normal fold change (FC(T/N)) of DLG2-AS1 expression in the 65 LUAD patients. Shown in a darker color are the patients who presented DLG2-AS1 downregulation (FC(T/N) < 0.66). *** = p -value < 0.001.
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    <t>ProBDNF</t> stably expressed in HEK293F cells is glycosylated. N-Glycosylation of proBDNF and the prodomain is documented by mass shift following deglycosylation with PNGase F: A, detection of proBDNF and BDNF by an antibody recognizing the BDNF region; B, detection of proBDNF and the prodomain by an antibody recognizing the prodomain region. The gel images were spliced as indicated by space to exclude samples not related to the study.
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    Alomone Labs uncleavable probdnf
    <t>ProBDNF</t> stably expressed in HEK293F cells is glycosylated. N-Glycosylation of proBDNF and the prodomain is documented by mass shift following deglycosylation with PNGase F: A, detection of proBDNF and BDNF by an antibody recognizing the BDNF region; B, detection of proBDNF and the prodomain by an antibody recognizing the prodomain region. The gel images were spliced as indicated by space to exclude samples not related to the study.
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    Alomone Labs mut probdnf
    <t>ProBDNF</t> stably expressed in HEK293F cells is glycosylated. N-Glycosylation of proBDNF and the prodomain is documented by mass shift following deglycosylation with PNGase F: A, detection of proBDNF and BDNF by an antibody recognizing the BDNF region; B, detection of proBDNF and the prodomain by an antibody recognizing the prodomain region. The gel images were spliced as indicated by space to exclude samples not related to the study.
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    Image Search Results


    LncRNA DLG2-AS1 is downregulated in lung adenocarcinoma (LUAD). ( a ) heatmap of the 90 lncRNAs analyzed with the Human LncRNA Profile Kit from System Biosciences (SBI); ( b ) DLG2-AS1 expression in normal and tumor samples; and ( c ) tumor/normal fold change (FC(T/N)) of DLG2-AS1 expression in the 65 LUAD patients. Shown in a darker color are the patients who presented DLG2-AS1 downregulation (FC(T/N) < 0.66). *** = p -value < 0.001.

    Journal: Cancers

    Article Title: LncRNA DLG2-AS1 as a Novel Biomarker in Lung Adenocarcinoma

    doi: 10.3390/cancers12082080

    Figure Lengend Snippet: LncRNA DLG2-AS1 is downregulated in lung adenocarcinoma (LUAD). ( a ) heatmap of the 90 lncRNAs analyzed with the Human LncRNA Profile Kit from System Biosciences (SBI); ( b ) DLG2-AS1 expression in normal and tumor samples; and ( c ) tumor/normal fold change (FC(T/N)) of DLG2-AS1 expression in the 65 LUAD patients. Shown in a darker color are the patients who presented DLG2-AS1 downregulation (FC(T/N) < 0.66). *** = p -value < 0.001.

    Article Snippet: Human DLG2 protein was detected using the anti-PSD-93 antibody (CAT# APZ-002, Alomone Labs, Jerusalem, Israel).

    Techniques: Expressing

    DLG2-AS1 restoration model on lung adenocarcinoma (LUAD) cell lines successfully overexpresses DLG2-AS1. DLG2-AS1 relative expression was measured by RT-qPCR 5 days after transfection, and normalized by the expression of the reference gene U1 snRNA. In all three cell lines tested, DLG2-AS1 was successfully overexpressed (relative expression (pLVX-DLG2AS/EV) > 2). * = p -value < 0.05.

    Journal: Cancers

    Article Title: LncRNA DLG2-AS1 as a Novel Biomarker in Lung Adenocarcinoma

    doi: 10.3390/cancers12082080

    Figure Lengend Snippet: DLG2-AS1 restoration model on lung adenocarcinoma (LUAD) cell lines successfully overexpresses DLG2-AS1. DLG2-AS1 relative expression was measured by RT-qPCR 5 days after transfection, and normalized by the expression of the reference gene U1 snRNA. In all three cell lines tested, DLG2-AS1 was successfully overexpressed (relative expression (pLVX-DLG2AS/EV) > 2). * = p -value < 0.05.

    Article Snippet: Human DLG2 protein was detected using the anti-PSD-93 antibody (CAT# APZ-002, Alomone Labs, Jerusalem, Israel).

    Techniques: Expressing, Quantitative RT-PCR, Transfection

    DLG2-AS1 is not a cis-regulator of DLG2 expression. ( a ) RNA expression levels of DLG2 were unaffected after DLG2-AS1 overexpression (pLVX-DLG2AS1) compared to empty vector transfection (EV). Results from RT-qPCR were normalized to expression of the reference gene U1 snRNA. ( b ) Protein levels of human DLG2 did not change upon transfection with pLVX-DLG2AS1 compared to EV in two independent Western blot experiments (relative pLVX-DLG2AS1/EV densitometry measure mean = 0.984 ± 0.199, p = 0.929). Uncropped WB images are available at the .

    Journal: Cancers

    Article Title: LncRNA DLG2-AS1 as a Novel Biomarker in Lung Adenocarcinoma

    doi: 10.3390/cancers12082080

    Figure Lengend Snippet: DLG2-AS1 is not a cis-regulator of DLG2 expression. ( a ) RNA expression levels of DLG2 were unaffected after DLG2-AS1 overexpression (pLVX-DLG2AS1) compared to empty vector transfection (EV). Results from RT-qPCR were normalized to expression of the reference gene U1 snRNA. ( b ) Protein levels of human DLG2 did not change upon transfection with pLVX-DLG2AS1 compared to EV in two independent Western blot experiments (relative pLVX-DLG2AS1/EV densitometry measure mean = 0.984 ± 0.199, p = 0.929). Uncropped WB images are available at the .

    Article Snippet: Human DLG2 protein was detected using the anti-PSD-93 antibody (CAT# APZ-002, Alomone Labs, Jerusalem, Israel).

    Techniques: Expressing, RNA Expression, Over Expression, Plasmid Preparation, Transfection, Quantitative RT-PCR, Western Blot

    DLG2-AS1 shows potential as a lung adenocarcinoma (LUAD) biomarker: ( a ) receiver operating characteristic (ROC) curve and area under curve (AUC) value of DLG2-AS1 expression in our cohort of LUAD patients; and ( b ) ROC curves and AUC values of other LUAD biomarkers (EGFR, TP53, MALAT-1, and NEAT1), obtained from TCGA data.

    Journal: Cancers

    Article Title: LncRNA DLG2-AS1 as a Novel Biomarker in Lung Adenocarcinoma

    doi: 10.3390/cancers12082080

    Figure Lengend Snippet: DLG2-AS1 shows potential as a lung adenocarcinoma (LUAD) biomarker: ( a ) receiver operating characteristic (ROC) curve and area under curve (AUC) value of DLG2-AS1 expression in our cohort of LUAD patients; and ( b ) ROC curves and AUC values of other LUAD biomarkers (EGFR, TP53, MALAT-1, and NEAT1), obtained from TCGA data.

    Article Snippet: Human DLG2 protein was detected using the anti-PSD-93 antibody (CAT# APZ-002, Alomone Labs, Jerusalem, Israel).

    Techniques: Biomarker Assay, Expressing

    Sequences of used oligonucleotides for qPCR and vector cloning.

    Journal: Cancers

    Article Title: LncRNA DLG2-AS1 as a Novel Biomarker in Lung Adenocarcinoma

    doi: 10.3390/cancers12082080

    Figure Lengend Snippet: Sequences of used oligonucleotides for qPCR and vector cloning.

    Article Snippet: Human DLG2 protein was detected using the anti-PSD-93 antibody (CAT# APZ-002, Alomone Labs, Jerusalem, Israel).

    Techniques: Plasmid Preparation, Clone Assay, Sequencing

    ProBDNF stably expressed in HEK293F cells is glycosylated. N-Glycosylation of proBDNF and the prodomain is documented by mass shift following deglycosylation with PNGase F: A, detection of proBDNF and BDNF by an antibody recognizing the BDNF region; B, detection of proBDNF and the prodomain by an antibody recognizing the prodomain region. The gel images were spliced as indicated by space to exclude samples not related to the study.

    Journal: The Journal of Biological Chemistry

    Article Title: N -Glycosylation is required for secretion of the precursor to brain-derived neurotrophic factor (proBDNF) carrying sulfated LacdiNAc structures

    doi: 10.1074/jbc.RA119.009989

    Figure Lengend Snippet: ProBDNF stably expressed in HEK293F cells is glycosylated. N-Glycosylation of proBDNF and the prodomain is documented by mass shift following deglycosylation with PNGase F: A, detection of proBDNF and BDNF by an antibody recognizing the BDNF region; B, detection of proBDNF and the prodomain by an antibody recognizing the prodomain region. The gel images were spliced as indicated by space to exclude samples not related to the study.

    Article Snippet: Glycosylated human proBDNF without C-terminal Myc-FLAG tags produced in HEK293F cells and nonglycosylated human proBDNF produced in E. coli (Alomone Labs, number B-257) were used to study the kinetic of cleavage by furin (R&D Systems, number 1503-SE-010).

    Techniques: Stable Transfection

    Extracted ion chromatograms (XIC) of (A) nonglycosylated, NYLDAANMSMR and (B) deglycosylated, NYLDAADMSMR glycopeptide of proBDNF secreted by HEK293F cells confirm site occupancy higher than 99%. C, tandem mass spectrum verifies deglycosylation of the proBDNF peptide under [18O]water.

    Journal: The Journal of Biological Chemistry

    Article Title: N -Glycosylation is required for secretion of the precursor to brain-derived neurotrophic factor (proBDNF) carrying sulfated LacdiNAc structures

    doi: 10.1074/jbc.RA119.009989

    Figure Lengend Snippet: Extracted ion chromatograms (XIC) of (A) nonglycosylated, NYLDAANMSMR and (B) deglycosylated, NYLDAADMSMR glycopeptide of proBDNF secreted by HEK293F cells confirm site occupancy higher than 99%. C, tandem mass spectrum verifies deglycosylation of the proBDNF peptide under [18O]water.

    Article Snippet: Glycosylated human proBDNF without C-terminal Myc-FLAG tags produced in HEK293F cells and nonglycosylated human proBDNF produced in E. coli (Alomone Labs, number B-257) were used to study the kinetic of cleavage by furin (R&D Systems, number 1503-SE-010).

    Techniques:

    Expression of wildtype (WT) proBDNF and its N121Q mutant (NQ) in stably transfected HEK293F cells. Protein expression was determined by Western blotting using an antibody, recognizing both BDNF and proBDNF, in the conditioned media (A) and cell lysates (B). Arrows point to (pro)BDNF forms: glycosylated proBDNF (black arrow), nonglycosylated proBDNF (white arrow), and mature BDNF (gray arrow). M indicates MagicMark molecular weight marker. Expression of mRNA was determined in WT and N121Q mutant by RT-PCR (C). ProBDNF/BDNF expression in cell lysates (D) or conditioned media (E) of HEK293F cells expressing N121Q mutant treated with proteasome inhibitor MG132 (MG), lysosomal inhibitor chloroquine (CQ), or chemical chaperone 4-phenylbutyric acid (PBA). The black arrow points to the position of nonglycosylated proBDNF.

    Journal: The Journal of Biological Chemistry

    Article Title: N -Glycosylation is required for secretion of the precursor to brain-derived neurotrophic factor (proBDNF) carrying sulfated LacdiNAc structures

    doi: 10.1074/jbc.RA119.009989

    Figure Lengend Snippet: Expression of wildtype (WT) proBDNF and its N121Q mutant (NQ) in stably transfected HEK293F cells. Protein expression was determined by Western blotting using an antibody, recognizing both BDNF and proBDNF, in the conditioned media (A) and cell lysates (B). Arrows point to (pro)BDNF forms: glycosylated proBDNF (black arrow), nonglycosylated proBDNF (white arrow), and mature BDNF (gray arrow). M indicates MagicMark molecular weight marker. Expression of mRNA was determined in WT and N121Q mutant by RT-PCR (C). ProBDNF/BDNF expression in cell lysates (D) or conditioned media (E) of HEK293F cells expressing N121Q mutant treated with proteasome inhibitor MG132 (MG), lysosomal inhibitor chloroquine (CQ), or chemical chaperone 4-phenylbutyric acid (PBA). The black arrow points to the position of nonglycosylated proBDNF.

    Article Snippet: Glycosylated human proBDNF without C-terminal Myc-FLAG tags produced in HEK293F cells and nonglycosylated human proBDNF produced in E. coli (Alomone Labs, number B-257) were used to study the kinetic of cleavage by furin (R&D Systems, number 1503-SE-010).

    Techniques: Expressing, Mutagenesis, Stable Transfection, Transfection, Western Blot, Molecular Weight, Marker, Reverse Transcription Polymerase Chain Reaction

    Kinetics of cleavage of glycosylated and nonglycosylated proBDNF by furin. Mass difference between glycosylated and nonglycosylated proBDNF enabled densitometric quantification of each proBDNF form: A, representative blot of the cleavage kinetic at the indicated time intervals; B, densitometric analysis of proBDNF intensities at the indicated time intervals. Results are expressed as percent of the proBDNF concentration (mean ± S.D., n = 3) at the beginning of the reaction. Note that proBDNF without C-terminal Myc-FLAG tag was used in the reaction as described under “Experimental procedures.”

    Journal: The Journal of Biological Chemistry

    Article Title: N -Glycosylation is required for secretion of the precursor to brain-derived neurotrophic factor (proBDNF) carrying sulfated LacdiNAc structures

    doi: 10.1074/jbc.RA119.009989

    Figure Lengend Snippet: Kinetics of cleavage of glycosylated and nonglycosylated proBDNF by furin. Mass difference between glycosylated and nonglycosylated proBDNF enabled densitometric quantification of each proBDNF form: A, representative blot of the cleavage kinetic at the indicated time intervals; B, densitometric analysis of proBDNF intensities at the indicated time intervals. Results are expressed as percent of the proBDNF concentration (mean ± S.D., n = 3) at the beginning of the reaction. Note that proBDNF without C-terminal Myc-FLAG tag was used in the reaction as described under “Experimental procedures.”

    Article Snippet: Glycosylated human proBDNF without C-terminal Myc-FLAG tags produced in HEK293F cells and nonglycosylated human proBDNF produced in E. coli (Alomone Labs, number B-257) were used to study the kinetic of cleavage by furin (R&D Systems, number 1503-SE-010).

    Techniques: Concentration Assay, FLAG-tag

    Mass spectra of proBDNF under the following conditions. A, precursor profile of the tryptic glycopeptide NYLDAANM(ox)SM(ox)R of proBDNF expressed in HEK293F cells; B, HCD fragmentation spectrum of the major glycoform (m/z 1132.42) at low collision energy (10% NCE) showing that characteristic LacdiNAc oxonium ion HexNAc-HexNAc (m/z 407.17) and its fucosylated form (m/z 553.22) with complementary y-ions (m/z 1373, 1381, 1454, and 1555) dominate the composition.

    Journal: The Journal of Biological Chemistry

    Article Title: N -Glycosylation is required for secretion of the precursor to brain-derived neurotrophic factor (proBDNF) carrying sulfated LacdiNAc structures

    doi: 10.1074/jbc.RA119.009989

    Figure Lengend Snippet: Mass spectra of proBDNF under the following conditions. A, precursor profile of the tryptic glycopeptide NYLDAANM(ox)SM(ox)R of proBDNF expressed in HEK293F cells; B, HCD fragmentation spectrum of the major glycoform (m/z 1132.42) at low collision energy (10% NCE) showing that characteristic LacdiNAc oxonium ion HexNAc-HexNAc (m/z 407.17) and its fucosylated form (m/z 553.22) with complementary y-ions (m/z 1373, 1381, 1454, and 1555) dominate the composition.

    Article Snippet: Glycosylated human proBDNF without C-terminal Myc-FLAG tags produced in HEK293F cells and nonglycosylated human proBDNF produced in E. coli (Alomone Labs, number B-257) were used to study the kinetic of cleavage by furin (R&D Systems, number 1503-SE-010).

    Techniques:

    Relative abundance of the glycoforms of HEK293F-produced proBDNF and prodomain expressed as % of all identified glycoforms

    Journal: The Journal of Biological Chemistry

    Article Title: N -Glycosylation is required for secretion of the precursor to brain-derived neurotrophic factor (proBDNF) carrying sulfated LacdiNAc structures

    doi: 10.1074/jbc.RA119.009989

    Figure Lengend Snippet: Relative abundance of the glycoforms of HEK293F-produced proBDNF and prodomain expressed as % of all identified glycoforms

    Article Snippet: Glycosylated human proBDNF without C-terminal Myc-FLAG tags produced in HEK293F cells and nonglycosylated human proBDNF produced in E. coli (Alomone Labs, number B-257) were used to study the kinetic of cleavage by furin (R&D Systems, number 1503-SE-010).

    Techniques: Produced

    Extracted ion chromatograms of four of the most abundant glycopeptides of proBDNF expressed in PC12 cells. A, C, and D, LacdiNAc containing structures; B, LacNAc containing structure. The arrow points to the peaks of glycopeptides aligned with slight retention time shifts as expected based on the hydrophilicity of the attached glycan. Numbers represent intensity of the extracted peaks (×105).

    Journal: The Journal of Biological Chemistry

    Article Title: N -Glycosylation is required for secretion of the precursor to brain-derived neurotrophic factor (proBDNF) carrying sulfated LacdiNAc structures

    doi: 10.1074/jbc.RA119.009989

    Figure Lengend Snippet: Extracted ion chromatograms of four of the most abundant glycopeptides of proBDNF expressed in PC12 cells. A, C, and D, LacdiNAc containing structures; B, LacNAc containing structure. The arrow points to the peaks of glycopeptides aligned with slight retention time shifts as expected based on the hydrophilicity of the attached glycan. Numbers represent intensity of the extracted peaks (×105).

    Article Snippet: Glycosylated human proBDNF without C-terminal Myc-FLAG tags produced in HEK293F cells and nonglycosylated human proBDNF produced in E. coli (Alomone Labs, number B-257) were used to study the kinetic of cleavage by furin (R&D Systems, number 1503-SE-010).

    Techniques:

    Impact of modulators of glycosylation on the BDNF/proBDNF ratio in HEK293F cells. Western blotting using antibodies to prodomain and mature BDNF regions was analyzed by densitometry for the following: A, intracellular BDNF to proBDNF ratio; B, secreted BDNF to proBDNF ratio; C, intracellular prodomain to proBDNF ratio; and D, secreted prodomain to proBDNF ratio. The representative Western blotting is labeled as the graphs: CTRL, control; CHLOR, 50 mm sodium chlorate; KIF, 1 μg/ml kifunensine; TNMC, 5 μg/ml of tunicamycin. Results are expressed as scatter plot (mean ± S.D., n = 3). *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared with CTRL, one-way analysis of variance with Bonferroni adjustment.

    Journal: The Journal of Biological Chemistry

    Article Title: N -Glycosylation is required for secretion of the precursor to brain-derived neurotrophic factor (proBDNF) carrying sulfated LacdiNAc structures

    doi: 10.1074/jbc.RA119.009989

    Figure Lengend Snippet: Impact of modulators of glycosylation on the BDNF/proBDNF ratio in HEK293F cells. Western blotting using antibodies to prodomain and mature BDNF regions was analyzed by densitometry for the following: A, intracellular BDNF to proBDNF ratio; B, secreted BDNF to proBDNF ratio; C, intracellular prodomain to proBDNF ratio; and D, secreted prodomain to proBDNF ratio. The representative Western blotting is labeled as the graphs: CTRL, control; CHLOR, 50 mm sodium chlorate; KIF, 1 μg/ml kifunensine; TNMC, 5 μg/ml of tunicamycin. Results are expressed as scatter plot (mean ± S.D., n = 3). *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared with CTRL, one-way analysis of variance with Bonferroni adjustment.

    Article Snippet: Glycosylated human proBDNF without C-terminal Myc-FLAG tags produced in HEK293F cells and nonglycosylated human proBDNF produced in E. coli (Alomone Labs, number B-257) were used to study the kinetic of cleavage by furin (R&D Systems, number 1503-SE-010).

    Techniques: Western Blot, Labeling