Structured Review

Selleck Chemicals leupeptin
PEDF induces degradation of the CMPK2 protein via a ubiquitin-dependent proteasomal mechanism. (A) Relative mRNA expression levels of CMPK2 in normal, LPS-induced, PEDF-overexpressing cells, and LPS + PEDF overexpression HTR8/SVneo cells; n=5. (B) Western blot and densitometry analysis of CMPK2 in HTR8/SVneo cells treated with LPS, <t>leupeptin</t> general protease inhibitor, NH4Cl lysosomal inhibitor, MG132 proteasome-specific inhibitor or DMSO for 6 h following overexpression of PEDF; n=5. (C) Cells were pre-treated with PEDF or MG132 and subsequently treated with LPS, and CMPK2 was immunoprecipitated; n=5. CMPK2 ubiquitination was determined using an anti-ubiquitin antibody. * P
Leupeptin, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 91 stars, based on 2 article reviews
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1) Product Images from "Pigment epithelium-derived factor facilitates NLRP3 inflammasome activation through downregulating cytidine monophosphate kinase 2: A potential treatment strategy for missed abortion"

Article Title: Pigment epithelium-derived factor facilitates NLRP3 inflammasome activation through downregulating cytidine monophosphate kinase 2: A potential treatment strategy for missed abortion

Journal: International Journal of Molecular Medicine

doi: 10.3892/ijmm.2020.4517

PEDF induces degradation of the CMPK2 protein via a ubiquitin-dependent proteasomal mechanism. (A) Relative mRNA expression levels of CMPK2 in normal, LPS-induced, PEDF-overexpressing cells, and LPS + PEDF overexpression HTR8/SVneo cells; n=5. (B) Western blot and densitometry analysis of CMPK2 in HTR8/SVneo cells treated with LPS, leupeptin general protease inhibitor, NH4Cl lysosomal inhibitor, MG132 proteasome-specific inhibitor or DMSO for 6 h following overexpression of PEDF; n=5. (C) Cells were pre-treated with PEDF or MG132 and subsequently treated with LPS, and CMPK2 was immunoprecipitated; n=5. CMPK2 ubiquitination was determined using an anti-ubiquitin antibody. * P
Figure Legend Snippet: PEDF induces degradation of the CMPK2 protein via a ubiquitin-dependent proteasomal mechanism. (A) Relative mRNA expression levels of CMPK2 in normal, LPS-induced, PEDF-overexpressing cells, and LPS + PEDF overexpression HTR8/SVneo cells; n=5. (B) Western blot and densitometry analysis of CMPK2 in HTR8/SVneo cells treated with LPS, leupeptin general protease inhibitor, NH4Cl lysosomal inhibitor, MG132 proteasome-specific inhibitor or DMSO for 6 h following overexpression of PEDF; n=5. (C) Cells were pre-treated with PEDF or MG132 and subsequently treated with LPS, and CMPK2 was immunoprecipitated; n=5. CMPK2 ubiquitination was determined using an anti-ubiquitin antibody. * P

Techniques Used: Expressing, Over Expression, Western Blot, Protease Inhibitor, Immunoprecipitation

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Multiple Displacement Amplification:

Article Title: Acetylation targets HSD17B4 for degradation via the CMA pathway in response to estrone
Article Snippet: .. HEK293T, MCF7 or MDA-MB-231 cells were lysed in 0.2 to 0.3% Nonidet P40 (Sigma-Aldrich, 74388) buffer containing 150 mM NaCl, 50 mM Tris-HCl, pH 7.5 and multiple protease inhibitors (PMSF [Ameresco, 0754; 1 mM], aprotinin [Selleckchem, S7377; 1 μg/ml], leupeptin [Selleckchem, S7380; 1 μg/ml], pepstatin [Sigma-Aldrich, T6522; 1 μg/ml], Na3 VO4 [Sigma-Aldrich, S6508; 1 mM], NaF [Sigma-Aldrich, S7920; 1 mM], TSA [1.5 mM], and NAM [15 mM], in final concentrations). .. Cell lysates were incubated for 3 h at 4°C with anti-FLAG M2 agarose (Sigma-Aldrich, A2220) after removal of debris by centrifuging at 4°C, 16,200 g for 12 to 15 min.

other:

Article Title: Acetylation targets HSD17B4 for degradation via the CMA pathway in response to estrone
Article Snippet: Proteasomal/lysosomal inhibitor treatments were performed by adding chloroquine (Sigma-Aldrich, C6628; 100 μΜ) for 8 h, MG132 (Abmole BioScience, M1902; 10 μM) for 6 h, leupeptin (Selleckchem, S7380; 0.4 mM) for 24 h, NH4 Cl (Sigma-Aldrich, A9434; 15 mM) for 24 h, wortmannin (Selleckchem, S2758; 100 nM) for 24 h, BAF (Selleckchem, S1413; 100 nM) for 8 h, respectively; all concentrations are final concentrations in the culture medium.

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    Selleck Chemicals leupeptin
    PEDF induces degradation of the CMPK2 protein via a ubiquitin-dependent proteasomal mechanism. (A) Relative mRNA expression levels of CMPK2 in normal, LPS-induced, PEDF-overexpressing cells, and LPS + PEDF overexpression HTR8/SVneo cells; n=5. (B) Western blot and densitometry analysis of CMPK2 in HTR8/SVneo cells treated with LPS, <t>leupeptin</t> general protease inhibitor, NH4Cl lysosomal inhibitor, MG132 proteasome-specific inhibitor or DMSO for 6 h following overexpression of PEDF; n=5. (C) Cells were pre-treated with PEDF or MG132 and subsequently treated with LPS, and CMPK2 was immunoprecipitated; n=5. CMPK2 ubiquitination was determined using an anti-ubiquitin antibody. * P
    Leupeptin, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/leupeptin/product/Selleck Chemicals
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    leupeptin - by Bioz Stars, 2020-07
    93/100 stars
      Buy from Supplier

    91
    Selleck Chemicals lysosome inhibitor leupeptin
    Nedd4-2 mediates the ubiquitination of glutamate transporter GLT-1 in MPP + -treated astrocytes. ( a ) Western blotting analysis showing that GLT-1 expression is decreased on MPP + treatment of astrocytes for 24 h in the membrane. ( b ) l -[ 3 H]-Glutamic acid uptake assay showing that 1 mM MPP + and 2 μ M PMA treatment for 24 h decreases the glutamate uptake in astrocytes. ( c and d ) Co-IP assays showing that GLT-1 is ubiquitinated after 24 h MPP + treatment of astrocytes. The capture antibody in c was anti-GLT-1; and in ( d ) was anti-Ub. IgG was tested as a negative control. ( e and f ) Co-IP assay showing the interaction between Nedd4-2 and GLT-1 in MPP + and PMA-treated astrocytes. The capture antibody in ( e ) was anti-Nedd4-2 and MPP + treatment time was 24 h; the capture antibody in ( f ) was anti-GLT-1 and MPP + treatment time was 24 h. IgG was tested as a negative control. ( g ) l -[ 3 H]-Glutamic acid uptake assay showing that lysosome inhibitor <t>(Leupeptin)</t> while not proteasome inhibitor (MG-132) increased glutamate uptake in MPP + -treated astrocytes for 24 h. ( h ) Western blotting analysis showing that Leupeptin while not MG-132 increased GLT-1 expression in MPP + -treated astrocytes for 24 h. Results are expressed as the mean±S.E. of at least three experiments. Student's t -test for Figures 1a, c and d , and One-way ANOVA for Figures 1b, e, f, g and h . *Represents a significant difference between other group and control group. ** P
    Lysosome Inhibitor Leupeptin, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lysosome inhibitor leupeptin/product/Selleck Chemicals
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    lysosome inhibitor leupeptin - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    Image Search Results


    PEDF induces degradation of the CMPK2 protein via a ubiquitin-dependent proteasomal mechanism. (A) Relative mRNA expression levels of CMPK2 in normal, LPS-induced, PEDF-overexpressing cells, and LPS + PEDF overexpression HTR8/SVneo cells; n=5. (B) Western blot and densitometry analysis of CMPK2 in HTR8/SVneo cells treated with LPS, leupeptin general protease inhibitor, NH4Cl lysosomal inhibitor, MG132 proteasome-specific inhibitor or DMSO for 6 h following overexpression of PEDF; n=5. (C) Cells were pre-treated with PEDF or MG132 and subsequently treated with LPS, and CMPK2 was immunoprecipitated; n=5. CMPK2 ubiquitination was determined using an anti-ubiquitin antibody. * P

    Journal: International Journal of Molecular Medicine

    Article Title: Pigment epithelium-derived factor facilitates NLRP3 inflammasome activation through downregulating cytidine monophosphate kinase 2: A potential treatment strategy for missed abortion

    doi: 10.3892/ijmm.2020.4517

    Figure Lengend Snippet: PEDF induces degradation of the CMPK2 protein via a ubiquitin-dependent proteasomal mechanism. (A) Relative mRNA expression levels of CMPK2 in normal, LPS-induced, PEDF-overexpressing cells, and LPS + PEDF overexpression HTR8/SVneo cells; n=5. (B) Western blot and densitometry analysis of CMPK2 in HTR8/SVneo cells treated with LPS, leupeptin general protease inhibitor, NH4Cl lysosomal inhibitor, MG132 proteasome-specific inhibitor or DMSO for 6 h following overexpression of PEDF; n=5. (C) Cells were pre-treated with PEDF or MG132 and subsequently treated with LPS, and CMPK2 was immunoprecipitated; n=5. CMPK2 ubiquitination was determined using an anti-ubiquitin antibody. * P

    Article Snippet: HTR8/SVneo cells were treated with LPS (10 μg/ml), leupeptin (5 μg/ml), NH4 Cl (10 μmol/l), MG132 (10 μmol/l), or DMSO (all purchased from Selleck Chemicals) for 6 h.

    Techniques: Expressing, Over Expression, Western Blot, Protease Inhibitor, Immunoprecipitation

    Nedd4-2 mediates the ubiquitination of glutamate transporter GLT-1 in MPP + -treated astrocytes. ( a ) Western blotting analysis showing that GLT-1 expression is decreased on MPP + treatment of astrocytes for 24 h in the membrane. ( b ) l -[ 3 H]-Glutamic acid uptake assay showing that 1 mM MPP + and 2 μ M PMA treatment for 24 h decreases the glutamate uptake in astrocytes. ( c and d ) Co-IP assays showing that GLT-1 is ubiquitinated after 24 h MPP + treatment of astrocytes. The capture antibody in c was anti-GLT-1; and in ( d ) was anti-Ub. IgG was tested as a negative control. ( e and f ) Co-IP assay showing the interaction between Nedd4-2 and GLT-1 in MPP + and PMA-treated astrocytes. The capture antibody in ( e ) was anti-Nedd4-2 and MPP + treatment time was 24 h; the capture antibody in ( f ) was anti-GLT-1 and MPP + treatment time was 24 h. IgG was tested as a negative control. ( g ) l -[ 3 H]-Glutamic acid uptake assay showing that lysosome inhibitor (Leupeptin) while not proteasome inhibitor (MG-132) increased glutamate uptake in MPP + -treated astrocytes for 24 h. ( h ) Western blotting analysis showing that Leupeptin while not MG-132 increased GLT-1 expression in MPP + -treated astrocytes for 24 h. Results are expressed as the mean±S.E. of at least three experiments. Student's t -test for Figures 1a, c and d , and One-way ANOVA for Figures 1b, e, f, g and h . *Represents a significant difference between other group and control group. ** P

    Journal: Cell Death & Disease

    Article Title: Regulation of glutamate transporter trafficking by Nedd4-2 in a Parkinson's disease model

    doi: 10.1038/cddis.2016.454

    Figure Lengend Snippet: Nedd4-2 mediates the ubiquitination of glutamate transporter GLT-1 in MPP + -treated astrocytes. ( a ) Western blotting analysis showing that GLT-1 expression is decreased on MPP + treatment of astrocytes for 24 h in the membrane. ( b ) l -[ 3 H]-Glutamic acid uptake assay showing that 1 mM MPP + and 2 μ M PMA treatment for 24 h decreases the glutamate uptake in astrocytes. ( c and d ) Co-IP assays showing that GLT-1 is ubiquitinated after 24 h MPP + treatment of astrocytes. The capture antibody in c was anti-GLT-1; and in ( d ) was anti-Ub. IgG was tested as a negative control. ( e and f ) Co-IP assay showing the interaction between Nedd4-2 and GLT-1 in MPP + and PMA-treated astrocytes. The capture antibody in ( e ) was anti-Nedd4-2 and MPP + treatment time was 24 h; the capture antibody in ( f ) was anti-GLT-1 and MPP + treatment time was 24 h. IgG was tested as a negative control. ( g ) l -[ 3 H]-Glutamic acid uptake assay showing that lysosome inhibitor (Leupeptin) while not proteasome inhibitor (MG-132) increased glutamate uptake in MPP + -treated astrocytes for 24 h. ( h ) Western blotting analysis showing that Leupeptin while not MG-132 increased GLT-1 expression in MPP + -treated astrocytes for 24 h. Results are expressed as the mean±S.E. of at least three experiments. Student's t -test for Figures 1a, c and d , and One-way ANOVA for Figures 1b, e, f, g and h . *Represents a significant difference between other group and control group. ** P

    Article Snippet: SGK inhibitor (GSK650394), PKC inhibitor (GF109203X), proteasome inhibitor (MG-132) and lysosome inhibitor (Leupeptin) were purchased from Selleck (Houston, TX, USA).

    Techniques: Western Blot, Expressing, Co-Immunoprecipitation Assay, Negative Control