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Monoclonal Antibodies

Anti-SUN2 [3.1E]

Invented at A*STAR Accelerate Technologies Pte Ltd
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Immunofluorescence analysis of paraffin-embedded mouse testis with 3.1E antibody. Testis from 5 months old C57BL/6 mice were embedded in paraffin and sectioned. Following heat-induced antigen retrieval (sodium citrate buffer pH 6) the paraffin sections were incubated with 1% Triton X-100 (in PBS) and subsequently stained with mouse monoclonal anti-Sun2 clone 3.1E antibody (red). Nuclei were visualized with Hoechst. In testis, Sun2 is specifically expressed in Sertoli cell and thus can be used as a marker for Sertoli cells.
Immunofluorescence staining of mouse tissues. Tissues from adult wild-type (upper two panels) or Sun2-null mice (lower panel) were cryoprotected in 30% sucrose before being transferred to OCT compound and frozen in liquid nitrogen. Tissue cryosections were fixed in 10% neutral buffered formalin and stained with Sun2 clone 3.1E antibody. Sun2 is highly expressed in ganglion cell layer of the retina (upper panel) and Sertoli cells (middle panel). Absence of 3.1E antibody staining in Sun2-null testis indicates that the antibody specifically recognises Sun2.
The 3.1E antibody specifically detects Sun2. Immortalised wild-type mouse adult fibroblast (MAFs Sun1/Sun2+/+) (upper panel), Sun2-null (MAFs Sun2-/-) (middle panel) and Sun1-null (MAFs Sun1-/-) (lower panel) were fixed in cold MeOH and labelled with the 3.1E monoclonal antibody. In Sun2-null but not Sun1-null MAFs there is a loss of 3.1E antibody staining signal indicating antibody specificity for Sun2.
Confocal immunofluorescence images of human HeLa cell line fixed with either 4% PFA (upper panel) or cold methanol (MeOH) (lower panel) and stained with 3.1E antibody. Nuclei were visualized with Hoechst.
Western blot analysis of 3.1E antibody. Whole cell lysates from mouse (C2C12, mouse adult fibroblasts (MAFs) and human (U2OS, 293T, Hela), cell lines were separated on SDS-PAGE gel, transferred to nitrocellulose membrane and blotted with anti-Sun2 of 3.1E antibody. Both primary and secondary antibodies were diluted in blocking buffer (5% nonfat dry milk re-suspended in 0.1% Tween and 1X PBS). The predicted molecular weight is 80.3kD and 81.6kDa for human and mouse Sun2 respectively. The observed band in Sun2-null MAFs most likely represent a truncated Sun2 cause by targeted exons deletion while generating Sun2 knockout mice.
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Catalogue Number 153484
Applications IF WB
Antigen/Gene or Protein Targets SUN-2
Synonyms FRIGG antibody, KIAA0668 antibody, Nuclear envelope protein antibody, Protein unc-84 homolog B antibody, Rab5 interacting protein antibody, RAB5IP antibody, Sad1 and UNC84 domain containing 2 antibody, Sad1 unc-84 domain protein 2 antibody, Sad1 unc84 domain protein antibody, Sad1/unc-84 protein-like 2 antibody, Sad1/unc84 protein-like antibody
Reactivity Mouse and Rat
Relevance A notable feature of metazoan cells is their morphological plasticity associated with acquisition of a range of complex activities. Implementation of numerous specialized functions during differentiation is dependent upon the rearrangement and transformation of organelles, including the cell nucleus. Crucial to many cell specific changes in nuclear localization and organization is establishment of connections between the nucleus and cytoskeleton. Such connections are formed in part by KASH-domain proteins of the outer nuclear membrane (ONM). In mammals six KASH proteins are known (Nesprins 1, 2, 3, 4, LRMP and KASH5). Appropriate localization of KASH proteins is dependent upon tethering in the ONM by SUN-domain proteins of the inner nuclear membrane. Together, SUN-KASH pairs form LINC complexes (LInker of the Nucleoskeleton and Cytoskeleton) that span both nuclear membranes and which couple nuclear structures to the cytoskeleton. In recent years it has become apparent that LINC complex defects may contribute to a number of human genetic diseases,
including muscular dystrophy, cerebellar ataxia, hearing loss and infertility.

In mammals the SUN protein family consists of at least 5 members. Sun1 and Sun2 share a high degree of homology and are ubiquitously expressed. Sun3-5 are more distantly related to Sun1-2 and have limited tissue expression.
Host Mouse
Immunogen Glutathione-S-transferase (GST) fused to residues 1-121 of human SUN2.
Immunogen UniProt ID Q9UH99
Positive Control Hela, C2C12, U2O2, fibroblasts
Subclass IgG2a kappa
Myeloma Used Sp2/0-Ag14
Strain C57BL/6
Notes Specificity: The antibody recognizes Sun2, it does not recognize Sun1
Epitope: Not known

Entrez Gene Number: 25777
Gene Symbol: FRIGG, KIAA0668, RAB5IP, UNC84B
UniProt Number: Q9UH99
Research Area Cell Signaling & Signal Transduction, Developmental Biology

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Invented at

A*STAR Accelerate Technologies Pte Ltd
A*STAR Accelerate Technologies Pte Ltd

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