Product nameAnti-NF2 / Merlin antibody [AF1G4]
See all NF2 / Merlin primary antibodies
DescriptionMouse monoclonal [AF1G4] to NF2 / Merlin
Tested applicationsSuitable for: ICC/IF, ChIP, WB, Indirect ELISA, IHC-Pmore details
Species reactivityReacts with: Mouse, Rat, Human
Recombinant human NF2/ Merlin with His tag purified from E.coli
- HeLa, 293T, MCF7, HepG2, MOLT4, C6 and L929 cell lysates. Human skeletal muscle FFPE tissue sections
This product was changed from ascites to tissue culture supernatant on 18th September 2017. Lot number GR310755 and higher lots are from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
Storage bufferPreservative: 0.03% Sodium azide
Constituents: HEPES, 0.01% BSA, 50% Glycerol, 0.87% Sodium chloride
PurityTissue culture supernatant
Light chain typekappa
Our Abpromise guarantee covers the use of ab88957 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|ChIP||Use at an assay dependent concentration.|
|WB||Use a concentration of 0.5 µg/ml. Detects a band of approximately 69 kDa (predicted molecular weight: 69 kDa).|
|Indirect ELISA||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
FunctionProbable regulator of the Hippo/SWH (Sav/Wts/Hpo) signaling pathway, a signaling pathway that plays a pivotal role in tumor suppression by restricting proliferation and promoting apoptosis. Along with WWC1 can synergistically induce the phosphorylation of LATS1 and LATS2 and can probably function in the regulation of the Hippo/SWH (Sav/Wts/Hpo) signaling pathway. May act as a membrane stabilizing protein. May inhibit PI3 kinase by binding to AGAP2 and impairing its stimulating activity. Suppresses cell proliferation and tumorigenesis by inhibiting the CUL4A-RBX1-DDB1-VprBP/DCAF1 E3 ubiquitin-protein ligase complex.
Tissue specificityWidely expressed. Isoform 1 and isoform 3 are predominant. Isoform 4, isoform 5 and isoform 6 are expressed moderately. Isoform 8 is found at low frequency. Isoform 7, isoform 9 and isoform 10 are not expressed in adult tissues, with the exception of adult retina expressing isoform 10. Isoform 9 is faintly expressed in fetal brain, heart, lung, skeletal muscle and spleen. Fetal thymus expresses isoforms 1, 7, 9 and 10 at similar levels.
Involvement in diseaseDefects in NF2 are the cause of neurofibromatosis 2 (NF2) [MIM:101000]; also known as central neurofibromatosis. NF2 is a genetic disorder characterized by bilateral vestibular schwannomas (formerly called acoustic neuromas), schwannomas of other cranial and peripheral nerves, meningiomas, and ependymomas. It is inherited in an autosomal dominant fashion with full penetrance. Affected individuals generally develop symptoms of eighth-nerve dysfunction in early adulthood, including deafness and balance disorder. Although the tumors of NF2 are histologically benign, their anatomic location makes management difficult, and patients suffer great morbidity and mortality.
Defects in NF2 are a cause of schwannomatosis (SCHWA) [MIM:162091]; also known as congenital cutaneous neurilemmomatosis. Schwannomas are benign tumors of the peripheral nerve sheath that usually occur singly in otherwise normal individuals. Multiple schwannomas in the same individual suggest an underlying tumor-predisposition syndrome. The most common such syndrome is NF2. The hallmark of NF2 is the development of bilateral vestibular-nerve schwannomas; but two-thirds or more of all NF2-affected individuals develop schwannomas in other locations, and dermal schwannomas may precede vestibular tumors in NF2-affected children. There have been several reports of individuals with multiple schwannomas who do not show evidence of vestibular schwannoma. Clinical report suggests that schwannomatosis is a clinical entity distinct from other forms of neurofibromatosis.
Sequence similaritiesContains 1 FERM domain.
modificationsPhosphorylation of Ser-518 inhibits nuclear localization by disrupting the intramolecular association of the FERM domain with the C-terminal tail.
Ubiquitinated by the CUL4A-RBX1-DDB1-DCAF1/VprBP E3 ubiquitin-protein ligase complex for ubiquitination and subsequent proteasome-dependent degradation.
Cellular localizationCytoplasm > perinuclear region. Cytoplasmic granule. Observed in cytoplasmic granules concentrated in a perinuclear location. Isoform 7 is absent from ruffling membranes and filopodia; Cytoplasm > perinuclear region. Cytoplasmic granule. Observed in cytoplasmic granules concentrated in a perinuclear location. Isoform 9 is absent from ruffling membranes and filopodia; Nucleus. Cell projection > filopodium membrane. Cell projection > ruffle membrane. Cytoplasm > perinuclear region. Cytoplasmic granule. Cytoplasm > cytoskeleton. In a fibroblastic cell line, isoform 10 is found homogeneously distributed over the entire cell, with a particularly strong staining in ruffling membranes and filopodia and Cell projection > filopodium membrane. Cell projection > ruffle membrane. Nucleus. In a fibroblastic cell line, isoform 1 is found homogeneously distributed over the entire cell, with a particularly strong staining in ruffling membranes and filopodia. Colocalizes with MPP1 in non-myelin-forming Schwann cells. Binds with VPRBP in the nucleus. The intramolecular association of the FERM domain with the C-terminal tail promotes nuclear accumulation. The unphosphorylated form accumulates predominantly in the nucleus while the phosphorylated form is largely confined to the non-nuclear fractions.
- Information by UniProt
- ACN antibody
- BANF antibody
- Bilateral acoustic neuroma antibody
Lane 1: Wild-type HAP1 cell lysate (40 µg)
Lane 2: Empty lane
Lane 3: NF2 / Merlin knockout HAP1 cell lysate (20 µg)
Lane 4: Empty lane
Lanes 1 - 4: Merged signal (red and green). Green - ab88957 observed at 70 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab88957 was shown to specifically react with NF2/Merlin in wild-type HAP1 cells. No band was observed when NF2 / Merlin knockout samples were examined. Wild-type and NF2/Merlin knockout samples were subjected to SDS-PAGE. Ab88957 and ab181602 (loading control to GAPDH) were diluted at 1/500 and 1/10,000 dilution respectively and incubated overnight at 4oC. Blots were developed with IRDye® 800CW Goat anti-Mouse IgG (H + L) ab216772 and IRDye® 680 Goat anti-Rabbit IgG (H + L) ab216777 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
IHC image of NF2/Merlin staining in human skeletal muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab88957, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ICC/IF image of ab88957 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab88957, 1/200 dilution) overnight at +4°C. The secondary antibody (green) was ab88957, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
All lanes : Anti-NF2 / Merlin antibody [AF1G4] (ab88957) at 1/2000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : 293T cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : HepG2 cell lysate
Lane 5 : MOLT4 cell lysate
Lane 6 : C6 cell lysate
Lane 7 : L929 cell lysate
Predicted band size: 69 kDa
Observed band size: 69 kDa
This product has been referenced in:
- Yoshida M et al. miR-7977 inhibits the Hippo-YAP signaling pathway in bone marrow mesenchymal stromal cells. PLoS One 14:e0213220 (2019). Read more (PubMed: 30835743) »
- Guo H et al. Determinants of lentiviral Vpx-CRL4 E3 ligase-mediated SAMHD1 degradation in the substrate adaptor protein DCAF1. Biochem Biophys Res Commun 513:933-939 (2019). Read more (PubMed: 31003777) »