Immunohistochemistry was performed on normal biopsies of deparaffinized Rat lymph node tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing Nuclear Pore-O-Linked Glycoprotein (MA1-071) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
|Tested species reactivity||Amphibian, Rat, Yeast|
|Published species reactivity||Yeast, Rat, Mouse, Human, Not Applicable|
|Host / Isotype||Mouse / IgM|
|Immunogen||Pore complex-lamina fraction purified from rat liver nuclear envelopes.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:1,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-071 detects nuclear pore-O-linked glycoprotein from rat, amphibian and yeast.
MA1-071 has been successfully used in Western blot, immunofluorescence and immunoprecipitation procedures. By Western blot, this antibody detects up to eight different proteins from the nuclear pore complex (NPC) of approximately 210, 180, 145, 100, 63, 58, 54 and 45 kDa. Immunofluorescence staining of NPC O-linked glycoproteins with MA1-071 results in exclusive labeling of the NPC proteins on a wide variety of mammalian cells as well as S. cerevisiae and Xenopus. Labeling occurs exclusively at the NPC with most of the labeling at the cytoplasmic and nucleoplasmic margins.
Microinjected MA1-071 inhibits both protein import and RNA export in Xenopus oocytes.
The MA1-071 immunogen is pore complex-lamina fraction purified from rat liver nuclear envelopes.
Diffusion of metabolites and small non-nuclear molecules as well as active, mediated import of protein and export of protein and RNA through the nuclear envelope occurs through nuclear pore complexes or NPC and quote;s. NPC and quote;s contain up to 100 different polypeptides which have a combined mass of about 125 megadaltons. The channel available for passive transport through the NPC is about 9-10 nm in diameter while carrier mediated changes in the NPC result in a ~25 nm channel used for larger, actively transported molecules. Of the 100 polypeptides, at least 8 of these are O-linked N-acetylglycosamine-modified in mammalian cells. All of the mammalian O-linked glycoproteins contain multiple copies of phenylalanine, glycine dipeptide repeats dispersed throughout part of their sequence. Studies indicate that the NPC O-linked glycoproteins have a direct role in nuclear protein import.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Cellular and Molecular Features of Developmentally Programmed Genome Rearrangement in a Vertebrate (Sea Lamprey: Petromyzon marinus).
MA1-071 was used in immunohistochemistry - paraffin section to study the sea lamprey (petromyzon marinus) vertebrate for molecular and cellular features of developmentally programmed genome rearrangement
|Timoshevskiy VA,Herdy JR,Keinath MC,Smith JJ||PLoS genetics (12:null)||2016|
Dynamic plasticity of large-scale chromatin structure revealed by self-assembly of engineered chromosome regions.
MA1-071 was used in immunocytochemistry to investigate the folding of engineered chromosome regions
|Sinclair P,Bian Q,Plutz M,Heard E,Belmont AS||The Journal of cell biology (190:761)||2010|
Lamin-binding fragment of LAP2 inhibits increase in nuclear volume during the cell cycle and progression into S phase.
MA1-071 was used in immunocytochemistry to investigate the functional properties of lamin-binding fragment of LAP2 during the cell cycle and progression into S phase
|Yang L,Guan T,Gerace L||The Journal of cell biology (139:1077)||1997|
A conditional allele of the novel repeat-containing yeast nucleoporin RAT7/NUP159 causes both rapid cessation of mRNA export and reversible clustering of nuclear pore complexes.
MA1-071 was used in immunocytochemistry to study how Rat7p regulates nucleocytoplasmic export of mRNA
|Gorsch LC,Dockendorff TC,Cole CN||The Journal of cell biology (129:939)||1995|
Nuclear protein import is inhibited by an antibody to a lumenal epitope of a nuclear pore complex glycoprotein.
MA1-071 was used in immunocytochemistry to investigate the role of gp210 in the assembly and functions of nuclear pore complex
|Greber UF,Gerace L||The Journal of cell biology (116:15)||1992|
Caenorhabditis elegans ortholog of a diabetes susceptibility locus: oga-1 (O-GlcNAcase) knockout impacts O-GlcNAc cycling, metabolism, and dauer.
MA1-071 was used in western blot to study the effect of O-GlcNAcase knockout in C. elegans.
|Forsythe ME,Love DC,Lazarus BD,Kim EJ,Prinz WA,Ashwell G,Krause MW,Hanover JA||Proceedings of the National Academy of Sciences of the United States of America (103:11952)||2006|
Exportin-5, a novel karyopherin, mediates nuclear export of double-stranded RNA binding proteins.
MA1-071 was used in western blot to identify a new karyopherin exportin-5 and its cellular function.
|Brownawell AM,Macara IG||The Journal of cell biology (156:53)||2002|
O-linked glycoproteins of the nuclear pore complex interact with a cytosolic factor required for nuclear protein import.
MA1-071 was used in western blot to characterize a cytosolic factor indispensable for nuclear protein import
|Sterne-Marr R,Blevitt JM,Gerace L||The Journal of cell biology (116:271)||1992|
Identification and characterization of Caenorhabditis elegans gamma-tubulin in dividing cells and differentiated tissues.
MA1-071 was used in immunohistochemistry to identify and characterize gamma-tubulin in Caenorhabditis elegans cells and tissues
|Bobinnec Y,Fukuda M,Nishida E||Journal of cell science (113 Pt 21:3747)||2000|
A role for RanBP1 in the release of CRM1 from the nuclear pore complex in a terminal step of nuclear export.
MA1-071 was used in immunoprecipitation to study the role of importin beta in nuclear export.
|Kehlenbach RH,Dickmanns A,Kehlenbach A,Guan T,Gerace L||The Journal of cell biology (145:645)||1999|