ccp1 buffer  (Millipore)


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    Structured Review

    Millipore ccp1 buffer
    Substrate specificity profile derived from putative <t>CCP1</t> substrates. ) was used to show the enriched residues present at the different identified CCP1 putative substrate positions as compared with the reference set ( i.e. C termini of human proteins in the Swiss-Prot database). For the iceLogo preparation we took into consideration that MCPs sequentially release amino acids. As a result, when a few amino acids are released to get to the final identified product, intermediate products of digestion act as new substrate and are further digested by CCP1. Consequently, intermediate products of digestion were considered as different individual substrates when preparing the iceLogo and, as a result, 35 CCP1 substrates were considered. Only those substrates identified by proteomics were considered here. In the representation the substrate residues are depicted according to Schechter Berger nomenclature. The frequency of the amino acid occurrence at each position in the sequence set was compared with the occurrence in the reference set. Only statistically significant residues with a p value ≤ 0.05 are plotted. Amino acids height shows the degree of difference in the positional amino acid frequencies in the experimental set as compared with the reference set. Residues that are statistically over- or underrepresented in the experimental set are shown in the upper or lower part of the iceLogo respectively.
    Ccp1 Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ccp1 buffer/product/Millipore
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ccp1 buffer - by Bioz Stars, 2020-03
    84/100 stars

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    affinity purified trad1

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    1) Product Images from "C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *"

    Article Title: C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *

    Journal: Molecular & Cellular Proteomics : MCP

    doi: 10.1074/mcp.M114.040360

    Substrate specificity profile derived from putative CCP1 substrates. ) was used to show the enriched residues present at the different identified CCP1 putative substrate positions as compared with the reference set ( i.e. C termini of human proteins in the Swiss-Prot database). For the iceLogo preparation we took into consideration that MCPs sequentially release amino acids. As a result, when a few amino acids are released to get to the final identified product, intermediate products of digestion act as new substrate and are further digested by CCP1. Consequently, intermediate products of digestion were considered as different individual substrates when preparing the iceLogo and, as a result, 35 CCP1 substrates were considered. Only those substrates identified by proteomics were considered here. In the representation the substrate residues are depicted according to Schechter Berger nomenclature. The frequency of the amino acid occurrence at each position in the sequence set was compared with the occurrence in the reference set. Only statistically significant residues with a p value ≤ 0.05 are plotted. Amino acids height shows the degree of difference in the positional amino acid frequencies in the experimental set as compared with the reference set. Residues that are statistically over- or underrepresented in the experimental set are shown in the upper or lower part of the iceLogo respectively.
    Figure Legend Snippet: Substrate specificity profile derived from putative CCP1 substrates. ) was used to show the enriched residues present at the different identified CCP1 putative substrate positions as compared with the reference set ( i.e. C termini of human proteins in the Swiss-Prot database). For the iceLogo preparation we took into consideration that MCPs sequentially release amino acids. As a result, when a few amino acids are released to get to the final identified product, intermediate products of digestion act as new substrate and are further digested by CCP1. Consequently, intermediate products of digestion were considered as different individual substrates when preparing the iceLogo and, as a result, 35 CCP1 substrates were considered. Only those substrates identified by proteomics were considered here. In the representation the substrate residues are depicted according to Schechter Berger nomenclature. The frequency of the amino acid occurrence at each position in the sequence set was compared with the occurrence in the reference set. Only statistically significant residues with a p value ≤ 0.05 are plotted. Amino acids height shows the degree of difference in the positional amino acid frequencies in the experimental set as compared with the reference set. Residues that are statistically over- or underrepresented in the experimental set are shown in the upper or lower part of the iceLogo respectively.

    Techniques Used: Derivative Assay, Activated Clotting Time Assay, Sequencing

    CCP1 overexpression increases the levels of Δ2-tubulin. A ). B , Detection of tubulin PTMs. Extracts from control cells or CCP1 expressing cells were analyzed by Western blot using anti-Δ2-tubulin and anti-Tyr-tubulin antibodies. The Western blot probing anti-β-actin served as loading control.
    Figure Legend Snippet: CCP1 overexpression increases the levels of Δ2-tubulin. A ). B , Detection of tubulin PTMs. Extracts from control cells or CCP1 expressing cells were analyzed by Western blot using anti-Δ2-tubulin and anti-Tyr-tubulin antibodies. The Western blot probing anti-β-actin served as loading control.

    Techniques Used: Over Expression, Expressing, Western Blot

    HMGB2 and HMGB1 are processed by CCP1. CCP1 is able to cleave the C terminus of A , HMGB1 and B , HMGB2. N-terminally Myc-tagged HMGB1 or HMGB2 were cotransfected with active or inactive CCP1. Equal amounts of each cell extract were analyzed by Western blot using a HA-tag antibody, to show equal expression levels of both CCP1 variants. Western blotting using an anti-Myc antibody shows the appearance of C-terminal processed forms of the HMGB proteins when co-expressed with active CCP1.
    Figure Legend Snippet: HMGB2 and HMGB1 are processed by CCP1. CCP1 is able to cleave the C terminus of A , HMGB1 and B , HMGB2. N-terminally Myc-tagged HMGB1 or HMGB2 were cotransfected with active or inactive CCP1. Equal amounts of each cell extract were analyzed by Western blot using a HA-tag antibody, to show equal expression levels of both CCP1 variants. Western blotting using an anti-Myc antibody shows the appearance of C-terminal processed forms of the HMGB proteins when co-expressed with active CCP1.

    Techniques Used: Western Blot, Expressing

    Related Articles

    High Performance Liquid Chromatography:

    Article Title: C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *
    Article Snippet: Affinity purified CCP1 was incubated at 5 ng/μl for 2 h or 16 h at 37 °C with affinity purified TRAD1 or HMGB3 at 25 ng/μl in CCP1 buffer (80 m m PIPES, pH 6.8, 1 m m MgCl2 , complemented with EDTA-free protease inhibitor mixture Set III (EMD Millipore)). .. Protein analysis of intact and processed HMGB3 was performed by nano LC-MS on a Waters nano-Acquity HPLC (Waters Corporation, Milford, MA) in-line coupled to an ESI-quadrupole (Q)-time-of-flight (TOF) (Waters Corporation) mass spectrometer.

    In Vitro:

    Article Title: C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *
    Article Snippet: Paragraph title: In Vitro Substrate Validation Assay ... Affinity purified CCP1 was incubated at 5 ng/μl for 2 h or 16 h at 37 °C with affinity purified TRAD1 or HMGB3 at 25 ng/μl in CCP1 buffer (80 m m PIPES, pH 6.8, 1 m m MgCl2 , complemented with EDTA-free protease inhibitor mixture Set III (EMD Millipore)).

    Liquid Chromatography with Mass Spectroscopy:

    Article Title: C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *
    Article Snippet: Affinity purified CCP1 was incubated at 5 ng/μl for 2 h or 16 h at 37 °C with affinity purified TRAD1 or HMGB3 at 25 ng/μl in CCP1 buffer (80 m m PIPES, pH 6.8, 1 m m MgCl2 , complemented with EDTA-free protease inhibitor mixture Set III (EMD Millipore)). .. Protein analysis of intact and processed HMGB3 was performed by nano LC-MS on a Waters nano-Acquity HPLC (Waters Corporation, Milford, MA) in-line coupled to an ESI-quadrupole (Q)-time-of-flight (TOF) (Waters Corporation) mass spectrometer.

    Protease Inhibitor:

    Article Title: C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *
    Article Snippet: .. Affinity purified CCP1 was incubated at 5 ng/μl for 2 h or 16 h at 37 °C with affinity purified TRAD1 or HMGB3 at 25 ng/μl in CCP1 buffer (80 m m PIPES, pH 6.8, 1 m m MgCl2 , complemented with EDTA-free protease inhibitor mixture Set III (EMD Millipore)). ..

    Incubation:

    Article Title: C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *
    Article Snippet: .. Affinity purified CCP1 was incubated at 5 ng/μl for 2 h or 16 h at 37 °C with affinity purified TRAD1 or HMGB3 at 25 ng/μl in CCP1 buffer (80 m m PIPES, pH 6.8, 1 m m MgCl2 , complemented with EDTA-free protease inhibitor mixture Set III (EMD Millipore)). ..

    Mass Spectrometry:

    Article Title: C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *
    Article Snippet: Affinity purified CCP1 was incubated at 5 ng/μl for 2 h or 16 h at 37 °C with affinity purified TRAD1 or HMGB3 at 25 ng/μl in CCP1 buffer (80 m m PIPES, pH 6.8, 1 m m MgCl2 , complemented with EDTA-free protease inhibitor mixture Set III (EMD Millipore)). .. Protein analysis of intact and processed HMGB3 was performed by nano LC-MS on a Waters nano-Acquity HPLC (Waters Corporation, Milford, MA) in-line coupled to an ESI-quadrupole (Q)-time-of-flight (TOF) (Waters Corporation) mass spectrometer.

    Affinity Purification:

    Article Title: C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *
    Article Snippet: .. Affinity purified CCP1 was incubated at 5 ng/μl for 2 h or 16 h at 37 °C with affinity purified TRAD1 or HMGB3 at 25 ng/μl in CCP1 buffer (80 m m PIPES, pH 6.8, 1 m m MgCl2 , complemented with EDTA-free protease inhibitor mixture Set III (EMD Millipore)). ..

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    Millipore ccp1 buffer
    Substrate specificity profile derived from putative <t>CCP1</t> substrates. ) was used to show the enriched residues present at the different identified CCP1 putative substrate positions as compared with the reference set ( i.e. C termini of human proteins in the Swiss-Prot database). For the iceLogo preparation we took into consideration that MCPs sequentially release amino acids. As a result, when a few amino acids are released to get to the final identified product, intermediate products of digestion act as new substrate and are further digested by CCP1. Consequently, intermediate products of digestion were considered as different individual substrates when preparing the iceLogo and, as a result, 35 CCP1 substrates were considered. Only those substrates identified by proteomics were considered here. In the representation the substrate residues are depicted according to Schechter Berger nomenclature. The frequency of the amino acid occurrence at each position in the sequence set was compared with the occurrence in the reference set. Only statistically significant residues with a p value ≤ 0.05 are plotted. Amino acids height shows the degree of difference in the positional amino acid frequencies in the experimental set as compared with the reference set. Residues that are statistically over- or underrepresented in the experimental set are shown in the upper or lower part of the iceLogo respectively.
    Ccp1 Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ccp1 buffer/product/Millipore
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ccp1 buffer - by Bioz Stars, 2020-03
    84/100 stars
      Buy from Supplier

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    Substrate specificity profile derived from putative CCP1 substrates. ) was used to show the enriched residues present at the different identified CCP1 putative substrate positions as compared with the reference set ( i.e. C termini of human proteins in the Swiss-Prot database). For the iceLogo preparation we took into consideration that MCPs sequentially release amino acids. As a result, when a few amino acids are released to get to the final identified product, intermediate products of digestion act as new substrate and are further digested by CCP1. Consequently, intermediate products of digestion were considered as different individual substrates when preparing the iceLogo and, as a result, 35 CCP1 substrates were considered. Only those substrates identified by proteomics were considered here. In the representation the substrate residues are depicted according to Schechter Berger nomenclature. The frequency of the amino acid occurrence at each position in the sequence set was compared with the occurrence in the reference set. Only statistically significant residues with a p value ≤ 0.05 are plotted. Amino acids height shows the degree of difference in the positional amino acid frequencies in the experimental set as compared with the reference set. Residues that are statistically over- or underrepresented in the experimental set are shown in the upper or lower part of the iceLogo respectively.

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *

    doi: 10.1074/mcp.M114.040360

    Figure Lengend Snippet: Substrate specificity profile derived from putative CCP1 substrates. ) was used to show the enriched residues present at the different identified CCP1 putative substrate positions as compared with the reference set ( i.e. C termini of human proteins in the Swiss-Prot database). For the iceLogo preparation we took into consideration that MCPs sequentially release amino acids. As a result, when a few amino acids are released to get to the final identified product, intermediate products of digestion act as new substrate and are further digested by CCP1. Consequently, intermediate products of digestion were considered as different individual substrates when preparing the iceLogo and, as a result, 35 CCP1 substrates were considered. Only those substrates identified by proteomics were considered here. In the representation the substrate residues are depicted according to Schechter Berger nomenclature. The frequency of the amino acid occurrence at each position in the sequence set was compared with the occurrence in the reference set. Only statistically significant residues with a p value ≤ 0.05 are plotted. Amino acids height shows the degree of difference in the positional amino acid frequencies in the experimental set as compared with the reference set. Residues that are statistically over- or underrepresented in the experimental set are shown in the upper or lower part of the iceLogo respectively.

    Article Snippet: Affinity purified CCP1 was incubated at 5 ng/μl for 2 h or 16 h at 37 °C with affinity purified TRAD1 or HMGB3 at 25 ng/μl in CCP1 buffer (80 m m PIPES, pH 6.8, 1 m m MgCl2 , complemented with EDTA-free protease inhibitor mixture Set III (EMD Millipore)).

    Techniques: Derivative Assay, Activated Clotting Time Assay, Sequencing

    CCP1 overexpression increases the levels of Δ2-tubulin. A ). B , Detection of tubulin PTMs. Extracts from control cells or CCP1 expressing cells were analyzed by Western blot using anti-Δ2-tubulin and anti-Tyr-tubulin antibodies. The Western blot probing anti-β-actin served as loading control.

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *

    doi: 10.1074/mcp.M114.040360

    Figure Lengend Snippet: CCP1 overexpression increases the levels of Δ2-tubulin. A ). B , Detection of tubulin PTMs. Extracts from control cells or CCP1 expressing cells were analyzed by Western blot using anti-Δ2-tubulin and anti-Tyr-tubulin antibodies. The Western blot probing anti-β-actin served as loading control.

    Article Snippet: Affinity purified CCP1 was incubated at 5 ng/μl for 2 h or 16 h at 37 °C with affinity purified TRAD1 or HMGB3 at 25 ng/μl in CCP1 buffer (80 m m PIPES, pH 6.8, 1 m m MgCl2 , complemented with EDTA-free protease inhibitor mixture Set III (EMD Millipore)).

    Techniques: Over Expression, Expressing, Western Blot

    HMGB2 and HMGB1 are processed by CCP1. CCP1 is able to cleave the C terminus of A , HMGB1 and B , HMGB2. N-terminally Myc-tagged HMGB1 or HMGB2 were cotransfected with active or inactive CCP1. Equal amounts of each cell extract were analyzed by Western blot using a HA-tag antibody, to show equal expression levels of both CCP1 variants. Western blotting using an anti-Myc antibody shows the appearance of C-terminal processed forms of the HMGB proteins when co-expressed with active CCP1.

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: C-terminomics Screen for Natural Substrates of Cytosolic Carboxypeptidase 1 Reveals Processing of Acidic Protein C termini *

    doi: 10.1074/mcp.M114.040360

    Figure Lengend Snippet: HMGB2 and HMGB1 are processed by CCP1. CCP1 is able to cleave the C terminus of A , HMGB1 and B , HMGB2. N-terminally Myc-tagged HMGB1 or HMGB2 were cotransfected with active or inactive CCP1. Equal amounts of each cell extract were analyzed by Western blot using a HA-tag antibody, to show equal expression levels of both CCP1 variants. Western blotting using an anti-Myc antibody shows the appearance of C-terminal processed forms of the HMGB proteins when co-expressed with active CCP1.

    Article Snippet: Affinity purified CCP1 was incubated at 5 ng/μl for 2 h or 16 h at 37 °C with affinity purified TRAD1 or HMGB3 at 25 ng/μl in CCP1 buffer (80 m m PIPES, pH 6.8, 1 m m MgCl2 , complemented with EDTA-free protease inhibitor mixture Set III (EMD Millipore)).

    Techniques: Western Blot, Expressing