Anti-MANF/ARMET-ATTO-594 Antibody

Mesencephalic astrocyte-derived neurotrophic factor, Arginine-rich mutated in early stage tumors, ARP
    Cat #: ANT-028-AR
    Alternative Name Mesencephalic astrocyte-derived neurotrophic factor, Arginine-rich mutated in early stage tumors, ARP
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Host: Rabbit
    Reactivity: m, r
    May also work in: h*
    Immunogen
    • Peptide (C)EVSKPLAHHIPVEK, corresponding to amino acid residues 91-104 of human MANF (Accession P55145). Mature MANF protein.
    • Anti-MANF/ARMET-ATTO-594 Antibody
    Accession (Uniprot) Number P55145
    Gene ID 7873
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Rat, mouse - identical.
    RRID AB_2756713.
    Purity Affinity purified on immobilized antigen.
    Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Isotype Rabbit IgG.
    Label ATTO-594. Maximum absorption 601 nm; Maximum fluorescence 627 nm. The fluorescence is excited most efficiently in the 580 – 615 nm range. This label belongs to the class of Rhodamine dyes and can be used with fluorescent equipment typically optimized to detect Texas Red and Alexa-594.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 50 µl double distilled water (DDW).
    Antibody concentration after reconstitution 1 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 × g 5 min).
    Negative control antigen storage before reconstitution Lyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
    Negative control antigen reconstitution 100 µl double distilled water (DDW).
    Negative control antigen storage after reconstitution -20°C.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis (unlabeled antibody, #ANT-028), and Immunohistochemical staining (labeled antibody).
    Applications: if, ih
    May also work in: ic*
    Immunohistochemistry
    • Anti-MANF/ARMET-ATTO-594 Antibody
      Expression of MANF in rat and mouse hippocampus
      Immunohistochemical staining of rat and mouse brain sections using Anti-MANF/ARMET-ATTO-594 Antibody (#ANT-028-AR), (1:80). A. MANF staining (red) in rat is detected in hippocampal dentate gyrus region in the cell soma (vertical arrows) and few astrocytes (horizontal arrow). B. MANF staining (red) is detected in same regions from mouse brain sections. Nuclei are stained using DAPI as the counterstain (blue).
    References
    1. Glembotski, C.C. (2011) J. Mol. Cell. Cardiol. 51, 512.
    2. Lindholm, P. and Saarma, M. (2010) Dev. Neurobiol. 70, 360.
    3. Parkash, V. et al. (2009) Protein Eng. Des. Sel. 22, 233.
    4. Airavaara, M. et al. (2012) Parkinsonism Relat. Disord. 18, 143.
    5. Lindholm, P. et al. (2008) Mol. Cell. Neurosci. 39, 356.
    6. Allen, S.J. et al. (2013) Pharmacol. Ther. 138, 155.
    7. Wang, H. et al. (2014) PLoS One 9, e90433.
    Scientific background

    The novel protein mesencephalic astrocyte-derived neurotrophic factor (MANF) is a survival molecule (alternatively referred to as arginine-rich, mutated in early stage tumors, or ARMET) that has high selectivity for dopaminergic neurons of the midbrain1,2. There, it induces proliferative and protective effects against neurodegeneration (usually caused by 6-hydroxydopamine); its therapeutic potential in dopamine-related neurodegenerative disorders such as Parkinson's disease is therefore promising3,4. MANF is also expressed cardiac tissues. As was recently suggested, it induces cardioprotection in the heart and anti-hypertrophic effects via reducing endoplasmic reticulum stress in ischemic conditions in both the heart and the brain – thus, its effects stretch beyond rescue and protection of dopaminergic neurons alone1,5.

    Generally, neurotrophic factors (NTFs) provide support, protection, and regulation for neurons – with each group of neurons having their own unique NTFs – both during development and at adulthood. Other noticeable NTFs include neurotrophin-3 and -4 (NT3 and NT4), nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), and glial cell derived neurotrophic factor (GDNF)6.

    MANF is considered to be evolutionary conserved within Mammalia and beyond (including distant species such as nematode and fruit fly – the latter shares more than 50% identity with human MANF gene) and is highly mutative in many tumors. It is mostly composed of alpha helices and random coils; its structure essentially differs from any other neurotrophic factor. It is suggested that the protein initiates intracellular signaling cascade by binding to cell-surface lipids and membranes via its cysteine-rich N-terminal. Together with its C-terminal which contains a disulfide bridge that endows a cytoprotective response against ER stress, MANF is implied to be bi-functional3,7.

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

    Species reactivity key:

    H- Human, M- Mouse, R- Rat
    Last update: 24/01/2020

    Anti-MANF/ARMET Antibody (#ANT-028) is a highly specific antibody directed against an epitope of the human protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize MANF from human, rat, and mouse samples.

    Anti-MANF/ARMET-ATTO-594 Antibody (#ANT-028-AR) is directly labeled with an ATTO-594 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-594 fluorescent label belongs to the class of Rhodamine dyes and can be used with fluorescent equipment typically optimized to detect Texas Red and Alexa-594. Anti-MANF/ARMET-ATTO-594 Antibody is especially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use
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