MABN1786

Anti-Merlin (NF2) Antibody, clone 1C4

clone 1C4, from mouse

• Sinónimos: Merlin, Moesin-ezrin-radixin-like protein, Neurofibromin-2, Schwannomerlin, Schwannomin
• UNSPSC Code: 12352203
• eCl@ss: 32160702

Propiedades

• biological source: mouse
• Quality Level: 100
• antibody form: purified antibody
• antibody product type: primary antibodies
• clone: 1C4, monoclonal
• species reactivity: rat, mouse, human
• technique(s): immunocytochemistry: suitable; immunoprecipitation (IP): suitable; western blot: suitable
• isotype: IgG1κ
• NCBI accession no.: NP_057502
• UniProt accession no.: P35240
• shipped in: ambient
• target post-translational modification: unmodified
• Gene Information: human ... NF2(4771)

Descripción

General description

Merlin (UniProt P35240; also known as Moesin-ezrin-radixin-like protein, Neurofibromin-2, Schwannomerlin, Schwannomin) is encoded by the NF2 (also known as SCH) gene (Gene ID 4771) in human. Originally identified as a tumor suppressor gene in the familial Neurofibromatosis type 2 (NF2) cancer patients, NF2 encodes the FERM (Ezrin, Radixin, Moesin) domain-containing merlin that plays a negative regulating role against cell growth and proliferation by functioning as an upstream activator of the Hippo pathway. Merlin directly activates Mst1/2 and recruits Lats1/2 to the plasma membrane for phosphorylation by Mst1/2. The phosphorylated Lats1/2 in turn inhibits YAP/TAZ transcription activity by phosphorylating YAP/TAZ and causes its cytoplasmic sequestration. Merlin also translocates to the nucleus, where it inhibits E3 ubiquitin ligase CRL4(DCAF1), thereby preventing it from ubiquitinating and downregulating Lats1/2. In its inactive state, the C-terminal tail domain (CTD; a.a. 507-595) of merlin is bound intramolecularly to its FERM domain (a.a. 19-313). Angiomotin (AMOT) induces merlin activation by binding to an elongated helical fragment within merlin CTD and subsequently releases the auto-inhibited conformation of Merlin. Merlin CTD can also undergo phosphorylation on Ser518 by p21-activated kinase (PAK) or cyclic AMP-dependent protein kinase (PKA), resulting in a weakened AMOT binding and a stabilization of merlin in its auto-inhibited state.

Specificity

Clone 1C4 detected merlin (NF2), but not any of the three ERM proteins, ezrin, radixin, and moesin (Gonzalez-Agosti, C., et al. (1996). Oncogene. 13(6):1239-1247). Clone 1C4 targeted epitope is present in human merlin (NF2) spliced isoforms 1 to 6, but not 7, 9, or 10 as reported by UniProt (P35240). Reactivity toward human spliced isoform 8 is possible, but has not been confirmed.

Immunogen

Recombinant protein corresponding to the C-terminal half of human merlin (NF2) isoform II (UniProt spliced isoform 3; P35240-3).

Application

Anti-Merlin (NF2), clone 1C4, Cat. No. MABN1786, is a highly specific mouse monoclonal antibody that targets Merlin (NF2) and has been tested in Immunocytochemistry, Immunoprecipitation, and Western Blotting.

Immunocytochemistry Analysis: A representative lot immunostained % paraformaldehyde-fixed, 0.2% Triton X-100-permeabilized wild-type, but not Nf2-knockout, mouse liver-derived epithelial cells (LDCs) by fluorescent immunocytochemistry. EGF-induced EGFR internalization was observed among Nf2-/-, but not Nf2+/+, LDCs (Curto, M., et al. (2007). J. Cell Biol. 177(5):893-903).

Immunocytochemistry Analysis: A representative lot detected merlin cellular distribution and HEI10 co-localization in a cell cycle-dependent manner by fluorescent immunocytochemistry staining of 3.5% paraformaldehyde-fixed U2OS human osteosarcoma cells (Grönholm, M., et al. (2006). Oncogene. 25(32):4389-4398).

Immunocytochemistry Analysis: A representative lot detected merlin cellular localization by fluorescent immunocytochemistry staining of 3.5% paraformaldehyde-fixed embryonic E16 rat neurons. Merlin was seen co-localized with RI to the cell body and extensions in a punctate pattern (Grönholm, M., et al. (2003). J. Biol. Chem. 278(42):41167-41172).

Immunocytochemistry Analysis: A representative lot detected merlin cellular localization by fluorescent immunocytochemistry staining of 4% paraformaldehyde-fixed, 0.1 NP-40-permeabilized human fibroblasts and primary meningioma cells. Merlin co-localized with F-actin at the leading and ruffling edges, but not at the stress fiber. No merlin co-localization with ezrin or moesin was observed (Gonzalez-Agosti, C., et al. (1996). Oncogene. 13(6):1239-1247).

Immunoprecipitation Analysis: A representative lot co-immunoprecipitated N-WASP with merlin from HEK293T cell lysate (Manchanda, N., et al. (2005). J. Biol. Chem. 280(13):12517-12522).

Western Blotting Analysis: A representative lot detected the presence of wild-type, but not L64P, merlin in the EGFR immunoprecipitates from Nf2-/- MEFs infected by adenovirus to express either wild-type or L64P merlin (Curto, M., et al. (2007). J. Cell Biol. 177(5):893-903).

Western Blotting Analysis: A representative lot detected a reduced merlin level in EGFR immunoprecipitate from mouse liver-derived epithelial cells (LDCs) upon shRNA-mediated NHE-RF1, but not NHE-RF2 knockdown. Merlin association with Ezrin was not affected by NHE-RF1 knockdown (Curto, M., et al. (2007). J. Cell Biol. 177(5):893-903).

Western Blotting Analysis: A representative lot detected a ~72 kDa merlin band in rat newborn fibroblast (RNF) and S-16 rat Schwann cell lysates, as well as a ~66 kDa merlin band in murine NIH/3T3 and human MRC-5 fetal fibroblast lystes without cross-reactivity toward the three ERM protein family members, ezrin, radixin, and moesin (Gonzalez-Agosti, C., et al. (1996). Oncogene. 13(6):1239-1247).

Research Category
Neuroscience

Quality

Evaluated by Western Blotting in HEK293 cell lysate.

Western Blotting Analysis: 4 µg/mL of this antibody detected Merlin (NF2) in 10 µg of HEK293 cell lysate.

Target description

~70 kDa observed. 69.78/68.71 kDa (mouse/rat), 69.69/72.51/69.09/59.10/64.19/64.00/85.35 kDa (human isoform 1/2/3/4/5/6/8) calculated. Human spliced isoform 3, 4, 5, and 6 are also known as isoform II, delE2/3, delE3, and delE2, respectively. Uncharacterized bands may be observed in some lysate(s).

Physical form

Format: Purified

Protein G purified

Purified mouse IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Información de seguridad

• Storage Class: 12 - Non Combustible Liquids
• wgk_germany: WGK 1
• flash_point_f: Not applicable
• flash_point_c: Not applicable