Human Metadherin Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human Metadherin in direct ELISAs and Western blots. In direct ELISAs, 100% cross-reactivity with recombinant mouse Metadherin is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human Metadherin
    Lys168-Ser298
    Accession # Q86UE4
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.2 µg/mL
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human Metadherin by Western Blot. Western blot shows lysates of MDA‑MB‑231 human breast cancer cell line, HeLa human cervical epithelial carcinoma cell line, and MDA‑MB‑453 human breast cancer cell line. PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human Metadherin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7644) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for Metadherin at approximately 80 and 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
Metadherin in HeLa Human Cell Line. Metadherin was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human Metadherin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7644) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.2 mg/mL.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Metadherin

MTDH (Metadherin; also LYRIC and Astrocyte Elevated Gene product-1/AEG1) is an 80 kDa membrane-bound protein that belongs to no known structural protein family. It has restricted expression, being found in breast ductal epithelium, striated muscle, select neurons and astrocytes, as well as multiple tumor types of diverse origin. MTDH is considered a mediator of Myc and Ha-Ras, and its activity is associated with stimulation of the beta -catenin/Wnt and NFkB signaling pathways. It is also believed to contribute to tight junction formation via an interaction with ZO-1. MTDH is also found in the nucleus where it complexes with CBP and p65. Human MTDH is 582 amino acids (aa) in length. It appears to be a type III transmembrane protein (N-terminus luminal/extracellular with no signal sequence) that is found embedded in both the plasma membrane and ER. It contains an N-terminal luminal region (aa 1-48), plus a large 513 aa cytoplasmic domain (aa 70-582) that possesses at least 11 utilized Ser/Thr phosphorylation sites. There is the potential for ubiquitination, SUMOylation, acetylation and proteolytic processing, and this may account for multiple bands in SDS-PAGE that represent MWs of 20, 35, 50-55, 70-75, and 86 kDa. Over aa 169-298, human MTDH shares 97% aa identity with mouse MDTH.

  • Entrez Gene IDs:
    92140 (Human); 67154 (Mouse); 170910 (Rat)
  • Alternate Names:
    3D3; AEG1; AEG-13D3/LYRIC; AEG1LYRIC/3D3; Astrocyte elevated gene-1 protein; LYRIC; LYRICMetastasis adhesion protein; Lysine-rich CEACAM1 co-isolated protein; Metadherin; MTDH; protein LYRIC
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