Actin Antibody

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Rufus W. Burlingame - One of the best experts on this subject based on the ideXlab platform.

  • Actin-reactive’ discriminated from ‘non-Actin-reactive’ smooth muscle autoAntibody by immunofluorescence reactivity with rat epithelial cell line
    Pathology, 2010
    Co-Authors: Roberta Taylor, Sharon Dearden, Cassandra C. Gill, Carol Buchner, Wendy Pollock, Rufus W. Burlingame
    Abstract:

    Aims: To compare smooth muscle Antibody (SMA) patterns in tissue sections with patterns in an immunofluorescence assay (IFA) using a rat intestinal epithelial cell line and results from an F-Actin IgG ELISA.Methods: SMA positive sera (n = 188) were classified by immunofluorescence staining of rodent kidney, stomach and liver sections as SMA-T (tubules) (n = 124) or SMA-V (vessels) (n = 64). The F-Actin pattern on the rat epithelial cell line was identified by immunofluorescence staining of Actin cables that was confirmed by dual immunofluorescence co-localisation with phalloidin.Results: Of 124 SMA-T positive sera, 123 reacted with the epithelial cell line and 120 with F-Actin by ELISA, giving sensitivity for detection of anti-F-Actin Antibody of 99% and 97%, respectively. Of 64 SMA-V positive sera, four reacted with the epithelial cell line (6%) and 41 with F-Actin by ELISA (64%). Tests of 493 normal blood donors and 100 disease controls yielded specificities of 584/593 (98.5%) and 562/593 (94.8%) for th...

  • A novel immunofluorescence test for f-Actin-reactive smooth muscle Antibody
    Pathology, 2010
    Co-Authors: Ban-hock Toh, Roberta Taylor, Sharon Dearden, Cassandra C. Gill, Carol Buchner, Wendy Pollock, Rufus W. Burlingame
    Abstract:

    Aim To compare smooth muscle Antibody (SMA) immunofluorescence staining patterns with an immunofluore-scence assay (IFA) using a rat intestinal epithelial cell line and with an F-Actin ELISA. Methods SMA sera (n = 188) were classified as SMA-T (n = 124) or SMA-V (n = 64) by immunofluorescence reactivity on composite rodent tissue sections of stomach, liver and kidney. Immunofluorsecence reactivity with the rat intestinal epithelial cell line was identified by staining of linear ‘Actin cables’ that was confirmed by dual immunof-luorescence co-localisation with F-Actin specifically labelled by phalloidin. Results Of 124 SMA-T positive sera, 123 reacted with the rat intestinal epithelial cell line and 120 with F-Actin by ELISA, giving sensitivities for detection of anti-F-Actin Antibody of 99% and 97%, respectively. In contrast, only four of 64 SMA-V positive sera reacted with the rat intestinal epithelial cell line (6%) while 41 reacted with F-Actin by ELISA (64%). Tests of 493 normal blood donors and 100 disease controls gave specificities of 584/593 (98.5%) and 561/593 (94.6%) for the cell line IFA and F-Actin ELISA respectively. Conclusions The rat intestinal epithelial cell line im-munofluorescence is a robust diagnostic assay for anti-F-Actin Antibody that can either replace the routine screening for anti-Actin SMA-T Antibody in tissue sections or be used as a confirmatory assay for anti-F-Actin Antibody after screening by F-Actin ELISA or by immunofluorescence on rodent tissue sections.

Peter E. Braun - One of the best experts on this subject based on the ideXlab platform.

  • 2′,3′‐Cyclic Nucleotide 3′‐Phosphodiesterase Binds to Actin‐Based Cytoskeletal Elements in an Isoprenylation‐Independent Manner
    Journal of Neurochemistry, 2002
    Co-Authors: Dino A. De Angelis, Peter E. Braun
    Abstract:

    : 2′,3′-Cyclic nucleotide 3′-phosphodiesterase (CNP) is an isoprenylated protein enriched in myelin and oligodendrocytes but also present in several other tissues at low levels. CNP binds avidly to membranes and in addition possesses several characteristics of cytoskeletal proteins. The role of isoprenylation in the association of CNP with the cytoskeleton was analyzed by ectopic expression in L cells of epitope-tagged CNP1 and a non-isoprenylated mutant CNP1. Using nonionic detergent extraction, drug-mediated cytoskeletal disruption, and coimmunoprecipitation with an anti-Actin Antibody, we show that CNP1 is associated with Actin-based cytoskeletal elements independently of its isoprenylation status. A control protein, p21c-H-ras, which is also modified by isoprenylation at its carboxyl-terminus, does not bind to cytoskeletal structures as judged by the same criteria. We present a model that accounts for the association of CNP1 with membranes and the cytoskeleton.

  • 2 3 cyclic nucleotide 3 phosphodiesterase binds to Actin based cytoskeletal elements in an isoprenylation independent manner
    Journal of Neurochemistry, 2002
    Co-Authors: Dino A. De Angelis, Peter E. Braun
    Abstract:

    : 2′,3′-Cyclic nucleotide 3′-phosphodiesterase (CNP) is an isoprenylated protein enriched in myelin and oligodendrocytes but also present in several other tissues at low levels. CNP binds avidly to membranes and in addition possesses several characteristics of cytoskeletal proteins. The role of isoprenylation in the association of CNP with the cytoskeleton was analyzed by ectopic expression in L cells of epitope-tagged CNP1 and a non-isoprenylated mutant CNP1. Using nonionic detergent extraction, drug-mediated cytoskeletal disruption, and coimmunoprecipitation with an anti-Actin Antibody, we show that CNP1 is associated with Actin-based cytoskeletal elements independently of its isoprenylation status. A control protein, p21c-H-ras, which is also modified by isoprenylation at its carboxyl-terminus, does not bind to cytoskeletal structures as judged by the same criteria. We present a model that accounts for the association of CNP1 with membranes and the cytoskeleton.

Yunching Huang - One of the best experts on this subject based on the ideXlab platform.

  • prominent expression of phosphodiesterase 5 in striated muscle of the rat urethra and levator ani
    The Journal of Urology, 2010
    Co-Authors: Yunching Huang, Guifang Wang
    Abstract:

    Purpose: We investigated phosphodiesterase 5 distribution and activity in the urethra.Materials and Methods: Rat tissues were examined for phosphodiesterase 5 and α-smooth muscle Actin expression. Urethral phosphodiesterase 5 activity was examined by tissue bath in the presence of sildenafil (Pfizer, New York, New York).Results: Anti-α-smooth muscle Actin Antibody (Abcam®) stained all known smooth muscles in all tested tissues and revealed a few smooth muscle fibers in the levator ani muscle. Anti-phosphodiesterase 5 Antibody (Abcam) stained smooth muscle in the penis and bladder but not striated leg muscle. However, it stained predominantly striated muscle in the urethra and the levator ani muscle. In the urethra the amount of phosphodiesterase 5 in striated muscle was 6 times that in smooth muscle. In urethral striated muscle phosphodiesterase 5 expression was localized to Z-band striations. Smooth and striated muscle intermingling was clearly visible on the inner and outer rims of the circularly arrang...

Francisco J B Sampaio - One of the best experts on this subject based on the ideXlab platform.

  • organization and relative content of smooth muscle cells collagen and elastic fibers in the corpus cavernosum of rat penis
    The Journal of Urology, 2000
    Co-Authors: Ana C A D Pinheiro, Waldemar S Costa, Luis Eduardo M Cardoso, Francisco J B Sampaio
    Abstract:

    Purpose: The corpus cavernosum smooth muscle and extracellular matrix are essential for normal penile erection and are implicated in erectile dysfunction. Although investigations of these issues have used the rat corpus cavernosum, organization of its components is to date not well known. We characterized and quantified the smooth muscle cells and the main extracellular matrix components of the rat corpus cavernosum.Materials and Methods: Collagen, elastic fibers and smooth muscle cells were stained on paraffin sections of rat penises using sirius red and Gomori’s reticulin, Weigert’s resorcin-fuchsin and an anti-smooth muscle cells α-Actin Antibody, respectively. Stained components were then quantified by computer aided morphometry.Results: Smooth muscle cells were restricted to the subendothelial space of corpus cavernosum and had a volumetric density of 9.1%. Collagen was thick, usually in transversely oriented bundles and was the most abundant component of the trabeculae with a volumetric density of 6...

Guifang Wang - One of the best experts on this subject based on the ideXlab platform.

  • prominent expression of phosphodiesterase 5 in striated muscle of the rat urethra and levator ani
    The Journal of Urology, 2010
    Co-Authors: Yunching Huang, Guifang Wang
    Abstract:

    Purpose: We investigated phosphodiesterase 5 distribution and activity in the urethra.Materials and Methods: Rat tissues were examined for phosphodiesterase 5 and α-smooth muscle Actin expression. Urethral phosphodiesterase 5 activity was examined by tissue bath in the presence of sildenafil (Pfizer, New York, New York).Results: Anti-α-smooth muscle Actin Antibody (Abcam®) stained all known smooth muscles in all tested tissues and revealed a few smooth muscle fibers in the levator ani muscle. Anti-phosphodiesterase 5 Antibody (Abcam) stained smooth muscle in the penis and bladder but not striated leg muscle. However, it stained predominantly striated muscle in the urethra and the levator ani muscle. In the urethra the amount of phosphodiesterase 5 in striated muscle was 6 times that in smooth muscle. In urethral striated muscle phosphodiesterase 5 expression was localized to Z-band striations. Smooth and striated muscle intermingling was clearly visible on the inner and outer rims of the circularly arrang...