A Monoclonal Antibody Toolkit for C. elegans

被引:65
作者
Hadwiger, Gayla [1 ]
Dour, Scott [1 ]
Arur, Swathi [2 ]
Fox, Paul [2 ]
Nonet, Michael L. [1 ]
机构
[1] Washington Univ, Sch Med, Dept Anat & Neurobiol, St Louis, MO 63110 USA
[2] Washington Univ, Sch Med, Dept Genet, St Louis, MO 63110 USA
来源
PLOS ONE | 2010年 / 5卷 / 04期
关键词
CELL-CYCLE PROGRESSION; CAENORHABDITIS-ELEGANS; NUCLEAR-LOCALIZATION; FUNCTIONAL-ANALYSIS; GENETIC-ANALYSIS; L1CAM HOMOLOG; IN-VIVO; PROTEIN; IDENTIFICATION; DYNAMIN;
D O I
10.1371/journal.pone.0010161
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background: Antibodies are critical tools in many avenues of biological research. Though antibodies can be produced in the research laboratory setting, most research labs working with vertebrates avail themselves of the wide array of commercially available reagents. By contrast, few such reagents are available for work with model organisms. Methodology/Principal Findings: We report the production of monoclonal antibodies directed against a wide range of proteins that label specific subcellular and cellular components, and macromolecular complexes. Antibodies were made to synaptobrevin (SNB-1), a component of synaptic vesicles; to Rim (UNC-10), a protein localized to synaptic active zones; to transforming acidic coiled-coil protein (TAC-1), a component of centrosomes; to CENP-C (HCP-4), which in worms labels the entire length of their holocentric chromosomes; to ORC2 (ORC-2), a subunit of the DNA origin replication complex; to the nucleolar phosphoprotein NOPP140 (DAO-5); to the nuclear envelope protein lamin (LMN-1); to EHD1 (RME-1) a marker for recycling endosomes; to caveolin (CAV-1), a marker for caveolae; to the cytochrome P450 (CYP-33E1), a resident of the endoplasmic reticulum; to beta-1,3-glucuronyltransferase (SQV-8) that labels the Golgi; to a chaperonin (HSP-60) targeted to mitochondria; to LAMP (LMP-1), a resident protein of lysosomes; to the alpha subunit of the 20S subcomplex (PAS-7) of the 26S proteasome; to dynamin (DYN-1) and to the alpha-subunit of the adaptor complex 2 (APA-2) as markers for sites of clathrin-mediated endocytosis; to the MAGUK, protein disks large (DLG-1) and cadherin (HMR-1), both of which label adherens junctions; to a cytoskeletal linker of the ezrin-radixin-moesin family (ERM-1), which localized to apical membranes; to an ERBIN family protein (LET-413) which localizes to the basolateral membrane of epithelial cells and to an adhesion molecule (SAX-7) which localizes to the plasma membrane at cell-cell contacts. In addition to working in whole mount immunocytochemistry, most of these antibodies work on western blots and thus should be of use for biochemical fractionation studies. Conclusions/Significance: We have produced a set of monoclonal antibodies to subcellular components of the nematode C. elegans for the research community. These reagents are being made available through the Developmental Studies Hybridoma Bank (DSHB).
引用
收藏
页数:14
相关论文
共 86 条
  • [1] Cellular and molecular determinants targeting the Caenorhabditis elegans PHR protein RPM-1 to perisynaptic regions
    Abrams, Benjamin
    Grill, Brock
    Huang, Xun
    Jin, Yishi
    [J]. DEVELOPMENTAL DYNAMICS, 2008, 237 (03) : 630 - 639
  • [2] PAR-3 is required for epithelial cell polarity in the distal spermatheca of C-elegans
    Aono, S
    Legouis, R
    Hoose, WA
    Kemphues, KJ
    [J]. DEVELOPMENT, 2004, 131 (12): : 2865 - 2874
  • [3] Multiple ERK substrates execute single biological processes in Caenorhabditis elegans germ-line development
    Arur, Swathi
    Ohmachi, Mitsue
    Nayak, Sudhir
    Hayes, Matthew
    Miranda, Alejandro
    Hay, Amanda
    Golden, Andy
    Schedl, Tim
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2009, 106 (12) : 4776 - 4781
  • [4] TAC-1 and ZYG-9 form a complex that promotes microtubule assembly in C-elegans embryos
    Bellanger, JM
    Gönczy, P
    [J]. CURRENT BIOLOGY, 2003, 13 (17) : 1488 - 1498
  • [5] Zonula adherens formation in Caenorhabditis elegans requires dlg-1, the homologue of the Drosophila gene discs large
    Bossinger, O
    Klebes, A
    Segbert, C
    Theres, C
    Knust, E
    [J]. DEVELOPMENTAL BIOLOGY, 2001, 230 (01) : 29 - 42
  • [6] The C-elegans hmr-1 gene can encode a neuronal classic cadherin involved in the regulation of axon fasciculation
    Broadbent, ID
    Pettitt, J
    [J]. CURRENT BIOLOGY, 2002, 12 (01) : 59 - 63
  • [7] Cell division -: A histone-H3-like protein in C-elegans
    Buchwitz, BJ
    Ahmad, K
    Moore, LL
    Roth, MB
    Henikoff, S
    [J]. NATURE, 1999, 401 (6753) : 547 - 548
  • [8] sqv-3 -7, and -8, a set of genes affecting morphogenesis in Caenorhabditis elegans, encode enzymes required for glycosaminoglycan biosynthesis
    Bulik, DA
    Wei, G
    Toyoda, H
    Kinoshita-Toyoda, A
    Waldrip, WR
    Esko, JD
    Robbins, PW
    Selleck, SB
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (20) : 10838 - 10843
  • [9] CAIRNS C, 1995, J CELL SCI, V108, P3339
  • [10] Development and evaluation of an in vivo assay in Caenorhabditis elegans for screening of compounds for their effect on cytochrome P450 expression
    Chakrapani, Baby P. S.
    Kumar, Sandeep
    Subramaniam, Jamuna R.
    [J]. JOURNAL OF BIOSCIENCES, 2008, 33 (02) : 269 - 277