sheep spinophilin antibody  (Thermo Fisher)


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    Structured Review

    Thermo Fisher sheep spinophilin antibody
    Validation of <t>spinophilin</t> interactions. WT or spinophilin KO striatal (STR) or olfactory tubercle (OT) lysates were immunoprecipitated with a spinophilin antibody. Lysates or immunoprecipitates were immunoblotted for spinophilin and three interacting proteins that were detected in the HA immunoprecipitates that had a decreased (SAP102) or increased (Clathrin heavy chain and SRCIN1) interaction with spinophilin in amphetamine-treated animals. Spinophilin and all associated proteins were detected in the spinophilin immunoprecipitates from WT animals, but were absent in immunoprecipitates isolated from KO animals.
    Sheep Spinophilin Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sheep spinophilin antibody/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sheep spinophilin antibody - by Bioz Stars, 2021-05
    86/100 stars

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    1) Product Images from "Proteomic Analysis of the Spinophilin Interactome in Rodent Striatum Following Psychostimulant Sensitization"

    Article Title: Proteomic Analysis of the Spinophilin Interactome in Rodent Striatum Following Psychostimulant Sensitization

    Journal: Proteomes

    doi: 10.3390/proteomes6040053

    Validation of spinophilin interactions. WT or spinophilin KO striatal (STR) or olfactory tubercle (OT) lysates were immunoprecipitated with a spinophilin antibody. Lysates or immunoprecipitates were immunoblotted for spinophilin and three interacting proteins that were detected in the HA immunoprecipitates that had a decreased (SAP102) or increased (Clathrin heavy chain and SRCIN1) interaction with spinophilin in amphetamine-treated animals. Spinophilin and all associated proteins were detected in the spinophilin immunoprecipitates from WT animals, but were absent in immunoprecipitates isolated from KO animals.
    Figure Legend Snippet: Validation of spinophilin interactions. WT or spinophilin KO striatal (STR) or olfactory tubercle (OT) lysates were immunoprecipitated with a spinophilin antibody. Lysates or immunoprecipitates were immunoblotted for spinophilin and three interacting proteins that were detected in the HA immunoprecipitates that had a decreased (SAP102) or increased (Clathrin heavy chain and SRCIN1) interaction with spinophilin in amphetamine-treated animals. Spinophilin and all associated proteins were detected in the spinophilin immunoprecipitates from WT animals, but were absent in immunoprecipitates isolated from KO animals.

    Techniques Used: Immunoprecipitation, Isolation

    Quantitation of spinophilin complexes isolated from dMSNs and iMSNs using tandem mass tag (TMT) analysis. ( A ) Striatal lysates isolated from male or female mice expressing HA spinophilin under the control of D1 or A2A promoters and treated with saline or amphetamine were immunoprecipitated with an HA antibody, digested with trypsin, labelled with eight different TMT tags, mixed and analyzed by mass spectrometry (MS/MS). ( B ) A higher intensity of TMT reporter abundance matching spinophilin was observed in amphetamine-treated compared to saline treated animals ( t -test; * p
    Figure Legend Snippet: Quantitation of spinophilin complexes isolated from dMSNs and iMSNs using tandem mass tag (TMT) analysis. ( A ) Striatal lysates isolated from male or female mice expressing HA spinophilin under the control of D1 or A2A promoters and treated with saline or amphetamine were immunoprecipitated with an HA antibody, digested with trypsin, labelled with eight different TMT tags, mixed and analyzed by mass spectrometry (MS/MS). ( B ) A higher intensity of TMT reporter abundance matching spinophilin was observed in amphetamine-treated compared to saline treated animals ( t -test; * p

    Techniques Used: Quantitation Assay, Isolation, Mouse Assay, Expressing, Immunoprecipitation, Mass Spectrometry

    Related Articles

    Transduction:

    Article Title: Selective targeting of the ?1 isoform of protein phosphatase 1 to F-actin in intact cells requires multiple domains in spinophilin and neurabin
    Article Snippet: .. Other antibodies were: mouse monoclonal PP2A (Transduction Laboratories), mouse monoclonal spinophilin (BD Pharmingen), mouse monoclonal neurabin (BD Pharmingen), mouse monoclonal GFP (Santa Cruz), mouse monoclonal myc (Zymed), donkey anti-mouse Cy5 (Jackson Immunoresearch), donkey anti-rabbit Alexa594 (Invitrogen), donkey anti-mouse Alexa647 (Invitrogen). .. A crude mixture of native protein phosphatase catalytic subunits was generated by ethanol precipitation of brain extracts, as previously described ( , ).

    Immunoprecipitation:

    Article Title: Proteomic Analysis of the Spinophilin Interactome in Rodent Striatum Following Psychostimulant Sensitization
    Article Snippet: 3 µg of goat HA polyclonal antibody (Bethyl Laboratories, Montgomery, TX, USA, A190-238A) or 5 µg goat spinophilin polyclonal antibody (Santa Cruz Biotechnology, Dallas, TX, USA, SC14774) were incubated at 4 °C with 750–800 µL (75–80%) of total striatal lysate overnight. .. Striatal cell lysates were immunoprecipitated with 1.6 µg of a sheep spinophilin antibody (ThermoFisher Scientific). .. The following day, protein G magnetic beads (DynaBeads, ThermoFisher Scientific) were added, and the mixture was incubated for 2 h. Beads were washed three times by magnetic separation in an immunoprecipitation wash buffer (50 mM NaCl, 50 mM Tris-HCl pH 7.5, 0.5% ( v / v ) Triton X-100).

    other:

    Article Title: Identification and Validation of Novel Spinophilin-associated Proteins in Rodent Striatum Using an Enhanced ex Vivo Shotgun Proteomics Approach *
    Article Snippet: The following antibodies were used as indicated: spinophilin: mouse monoclonal antibody (BD Biosciences 612166; epitope mapping between rat spinophilin residues 238–348), rabbit polyclonal spinophilin antibody (Millipore 06-852; epitope mapping between rat spinophilin residues 286–390), and goat polyclonal antibody (Santa Cruz Biotechnology SC-14774; epitope mapping to residues ∼50–100 of rat spinophilin); CaMKII: goat polyclonal antibody , mouse monoclonal CaMKIIα antibody (Affinity Bioreagents/Thermo Fisher Scientific MA1-048), and mouse monoclonal CaMKIIβ antibody (Zymed Laboratories Inc./Invitrogen 13-9800); pan-α-actinin, rabbit polyclonal antibody (Santa Cruz Biotechnology SC-15335); densin: goat polyclonal antibodies 450 and 650 ( ) and rabbit polyclonal antibody BΔN ( ); thousand and one amino acid kinase 1 (TAO1): rabbit polyclonal antibody (Bethyl Laboratories A300-524A); TAO3: rabbit polyclonal antibody (Bethyl Laboratories A300-536A); Tiam1: rabbit polyclonal antibody (Santa Cruz Biotechnology SC-872); P70S6 kinase: rabbit polyclonal antibody (Santa Cruz Biotechnology SC-230); doublecortin: rabbit polyclonal antibody (Cell Signaling Technology 4604); RasGrf1: rabbit polyclonal antibody (Santa Cruz Biotechnology SC-224); PP1γ1: sheep polyclonal antibody ( ); PSD-95 (NeuroMab 75-028); tyrosine hydroxylase (ImmunoStar 22941); neurabin: mouse monoclonal antibody (BD Transduction Laboratories 611088); myosin Va: rabbit polyclonal antibody (Sigma M4812); neurofilament light polypeptide: rabbit (Cell Signaling Technology 2837) and mouse (Santa Cruz Biotechnology SC-58559) antibodies; Myc: mouse monoclonal antibody (Vanderbilt Monoclonal Antibody Core); HA: mouse monoclonal antibody (Vanderbilt Monoclonal Antibody Core) and rabbit polyclonal antibody (Santa Cruz Biotechnology SC-805); and fluorescent secondary antibodies: donkey anti-mouse Alexa Fluor 546, donkey anti-rabbit Alexa Fluor 488, and donkey anti-goat Alexa Fluor 633 (Molecular Probes).

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    Thermo Fisher sheep spinophilin antibody
    Generation and characterization of Cre-expressing, HA-tagged human <t>spinophilin</t> mice. ( A ) A construct containing DNA encoding HA-tagged human spinophilin with a P2A sequence and mNeptune3 fluorescent protein was cloned into the pBIGT vector that contains a floxed-stop sequence. ( B ) The construct encoding the floxed-stop sequence and the HA-spinophilin-P2A-mNeptune 3 sequence was subcloned into the ROSA targeting vector pROSA_26.PA. ( C ) The modified ROSA vector was used for generation of the targeted transgenic mice. ( D ) Striatal cells were transfected without or with HA-tagged human spinophilin. Lysates were immunoprecipitated with either an HA or spinophilin antibody and immunoblotted with an HA antibody or a spinophilin antibody. HA-spinophilin was selectively detected when it was overexpressed. ( E ) Mice express HA-tagged spinophilin upon crossing with Cre recombinase expressed in the direct pathway (D1) or indirect pathway (A2A) medium spiny neurons. ( F ) Spinophilin and protein phosphatase 1 immunoblots of inputs and HA-immunoprecipitates from HA spinophilin mice crossed with D1 or A2A Cre-recombinase-expressing mice. ( G ) Mice expressing HA-spinophilin had non-significant increases in total spinophilin expression.
    Sheep Spinophilin Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sheep spinophilin antibody/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sheep spinophilin antibody - by Bioz Stars, 2021-05
    97/100 stars
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    Generation and characterization of Cre-expressing, HA-tagged human spinophilin mice. ( A ) A construct containing DNA encoding HA-tagged human spinophilin with a P2A sequence and mNeptune3 fluorescent protein was cloned into the pBIGT vector that contains a floxed-stop sequence. ( B ) The construct encoding the floxed-stop sequence and the HA-spinophilin-P2A-mNeptune 3 sequence was subcloned into the ROSA targeting vector pROSA_26.PA. ( C ) The modified ROSA vector was used for generation of the targeted transgenic mice. ( D ) Striatal cells were transfected without or with HA-tagged human spinophilin. Lysates were immunoprecipitated with either an HA or spinophilin antibody and immunoblotted with an HA antibody or a spinophilin antibody. HA-spinophilin was selectively detected when it was overexpressed. ( E ) Mice express HA-tagged spinophilin upon crossing with Cre recombinase expressed in the direct pathway (D1) or indirect pathway (A2A) medium spiny neurons. ( F ) Spinophilin and protein phosphatase 1 immunoblots of inputs and HA-immunoprecipitates from HA spinophilin mice crossed with D1 or A2A Cre-recombinase-expressing mice. ( G ) Mice expressing HA-spinophilin had non-significant increases in total spinophilin expression.

    Journal: Proteomes

    Article Title: Proteomic Analysis of the Spinophilin Interactome in Rodent Striatum Following Psychostimulant Sensitization

    doi: 10.3390/proteomes6040053

    Figure Lengend Snippet: Generation and characterization of Cre-expressing, HA-tagged human spinophilin mice. ( A ) A construct containing DNA encoding HA-tagged human spinophilin with a P2A sequence and mNeptune3 fluorescent protein was cloned into the pBIGT vector that contains a floxed-stop sequence. ( B ) The construct encoding the floxed-stop sequence and the HA-spinophilin-P2A-mNeptune 3 sequence was subcloned into the ROSA targeting vector pROSA_26.PA. ( C ) The modified ROSA vector was used for generation of the targeted transgenic mice. ( D ) Striatal cells were transfected without or with HA-tagged human spinophilin. Lysates were immunoprecipitated with either an HA or spinophilin antibody and immunoblotted with an HA antibody or a spinophilin antibody. HA-spinophilin was selectively detected when it was overexpressed. ( E ) Mice express HA-tagged spinophilin upon crossing with Cre recombinase expressed in the direct pathway (D1) or indirect pathway (A2A) medium spiny neurons. ( F ) Spinophilin and protein phosphatase 1 immunoblots of inputs and HA-immunoprecipitates from HA spinophilin mice crossed with D1 or A2A Cre-recombinase-expressing mice. ( G ) Mice expressing HA-spinophilin had non-significant increases in total spinophilin expression.

    Article Snippet: Striatal cell lysates were immunoprecipitated with 1.6 µg of a sheep spinophilin antibody (ThermoFisher Scientific).

    Techniques: Expressing, Mouse Assay, Construct, Sequencing, Clone Assay, Plasmid Preparation, Modification, Transgenic Assay, Transfection, Immunoprecipitation, Western Blot

    Validation of spinophilin interactions. WT or spinophilin KO striatal (STR) or olfactory tubercle (OT) lysates were immunoprecipitated with a spinophilin antibody. Lysates or immunoprecipitates were immunoblotted for spinophilin and three interacting proteins that were detected in the HA immunoprecipitates that had a decreased (SAP102) or increased (Clathrin heavy chain and SRCIN1) interaction with spinophilin in amphetamine-treated animals. Spinophilin and all associated proteins were detected in the spinophilin immunoprecipitates from WT animals, but were absent in immunoprecipitates isolated from KO animals.

    Journal: Proteomes

    Article Title: Proteomic Analysis of the Spinophilin Interactome in Rodent Striatum Following Psychostimulant Sensitization

    doi: 10.3390/proteomes6040053

    Figure Lengend Snippet: Validation of spinophilin interactions. WT or spinophilin KO striatal (STR) or olfactory tubercle (OT) lysates were immunoprecipitated with a spinophilin antibody. Lysates or immunoprecipitates were immunoblotted for spinophilin and three interacting proteins that were detected in the HA immunoprecipitates that had a decreased (SAP102) or increased (Clathrin heavy chain and SRCIN1) interaction with spinophilin in amphetamine-treated animals. Spinophilin and all associated proteins were detected in the spinophilin immunoprecipitates from WT animals, but were absent in immunoprecipitates isolated from KO animals.

    Article Snippet: Striatal cell lysates were immunoprecipitated with 1.6 µg of a sheep spinophilin antibody (ThermoFisher Scientific).

    Techniques: Immunoprecipitation, Isolation

    Greater abundance of spinophilin interacting proteins in amphetamine-treated animals occurs across both sexes and cell types. ( A ) Principal component analysis of individual samples normalized to spinophilin abundance within each sample and filtered for eight or more PSMs. ( B ) A volcano plot showing a majority of the proteins have increased abundance in HA immunoprecipitates isolated from amphetamine treated animals when normalized to the amphetamine-dependent increase in spinophilin abundance. ( C ) A plot of the abundance of spinophilin interacting proteins isolated from male, D1 Cre expressing animals and normalized for spinophilin expression ( Table S2 ) and quantified from treated (Y-axis) or control (X-axis) samples. Left panel shows mean ± standard deviation, the right panel just shows the mean of the two values. ( D ) A plot of the abundance of spinophilin interacting proteins isolated from female, D1 Cre expressing animals and normalized for spinophilin expression ( Table S2 ) and quantified from treated (Y-axis) or control (X-axis) samples. ( E ) A plot of the abundance of spinophilin interacting proteins isolated from female, A2A-Cre expressing animals and normalized for spinophilin expression ( Table S2 ) and quantified from treated (Y-axis) or control (X-axis) samples.

    Journal: Proteomes

    Article Title: Proteomic Analysis of the Spinophilin Interactome in Rodent Striatum Following Psychostimulant Sensitization

    doi: 10.3390/proteomes6040053

    Figure Lengend Snippet: Greater abundance of spinophilin interacting proteins in amphetamine-treated animals occurs across both sexes and cell types. ( A ) Principal component analysis of individual samples normalized to spinophilin abundance within each sample and filtered for eight or more PSMs. ( B ) A volcano plot showing a majority of the proteins have increased abundance in HA immunoprecipitates isolated from amphetamine treated animals when normalized to the amphetamine-dependent increase in spinophilin abundance. ( C ) A plot of the abundance of spinophilin interacting proteins isolated from male, D1 Cre expressing animals and normalized for spinophilin expression ( Table S2 ) and quantified from treated (Y-axis) or control (X-axis) samples. Left panel shows mean ± standard deviation, the right panel just shows the mean of the two values. ( D ) A plot of the abundance of spinophilin interacting proteins isolated from female, D1 Cre expressing animals and normalized for spinophilin expression ( Table S2 ) and quantified from treated (Y-axis) or control (X-axis) samples. ( E ) A plot of the abundance of spinophilin interacting proteins isolated from female, A2A-Cre expressing animals and normalized for spinophilin expression ( Table S2 ) and quantified from treated (Y-axis) or control (X-axis) samples.

    Article Snippet: Striatal cell lysates were immunoprecipitated with 1.6 µg of a sheep spinophilin antibody (ThermoFisher Scientific).

    Techniques: Isolation, Expressing, Standard Deviation

    Quantitation of spinophilin complexes isolated from dMSNs and iMSNs using tandem mass tag (TMT) analysis. ( A ) Striatal lysates isolated from male or female mice expressing HA spinophilin under the control of D1 or A2A promoters and treated with saline or amphetamine were immunoprecipitated with an HA antibody, digested with trypsin, labelled with eight different TMT tags, mixed and analyzed by mass spectrometry (MS/MS). ( B ) A higher intensity of TMT reporter abundance matching spinophilin was observed in amphetamine-treated compared to saline treated animals ( t -test; * p

    Journal: Proteomes

    Article Title: Proteomic Analysis of the Spinophilin Interactome in Rodent Striatum Following Psychostimulant Sensitization

    doi: 10.3390/proteomes6040053

    Figure Lengend Snippet: Quantitation of spinophilin complexes isolated from dMSNs and iMSNs using tandem mass tag (TMT) analysis. ( A ) Striatal lysates isolated from male or female mice expressing HA spinophilin under the control of D1 or A2A promoters and treated with saline or amphetamine were immunoprecipitated with an HA antibody, digested with trypsin, labelled with eight different TMT tags, mixed and analyzed by mass spectrometry (MS/MS). ( B ) A higher intensity of TMT reporter abundance matching spinophilin was observed in amphetamine-treated compared to saline treated animals ( t -test; * p

    Article Snippet: Striatal cell lysates were immunoprecipitated with 1.6 µg of a sheep spinophilin antibody (ThermoFisher Scientific).

    Techniques: Quantitation Assay, Isolation, Mouse Assay, Expressing, Immunoprecipitation, Mass Spectrometry