Human Melanotransferrin/CD228 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human Melanotransferrin/CD228 in ELISAs.
  • Source
    Monoclonal Mouse IgG1 Clone # 893438
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    HEK293 human embryonic kidney cell line transfected with human Melanotransferrin/CD228
    Met1-Gly711
    Accession # P08582
  • 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
    1 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
  • Immunohistochemistry
    5-25 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 Melanotransferrin/CD228 by Western Blot.

Western blot shows lysates of SK‑Mel‑28 human malignant melanoma cell line and Hs 294T human melanoma cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Melanotransferrin/CD228 Monoclonal Antibody (Catalog # MAB8175) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Melanotransferrin/CD228 at approximately 90-95 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry
Melanotransferrin/CD228 in Human Melanoma Tissue. Melanotransferrin/CD228 was detected in perfusion fixed frozen sections of human melanoma tissue using Mouse Anti-Human Melanotransferrin/CD228 Monoclonal Antibody (Catalog # MAB8175) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane and cytoplasm. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Detection of Human Melanotransferrin/CD228 by Simple WesternTM. Simple Western lane view shows lysates of SK‑Mel‑28 human malignant melanoma cell line, loaded at 0.5 mg/mL. A specific band was detected for Melanotransferrin/CD228 at approximately 109 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human Melanotransferrin/CD228 Monoclonal Antibody (Catalog # MAB8175). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • 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: Melanotransferrin/CD228
Melanotransferrin, also known as MTF, CD228, melanoma-associated antigen p97, MAP97 and MFI2, is a 90-97 kDa sialoglycoprotein member of the transferrin family. Unlike other transferrins, which are secreted, MTF is usually found tethered to the cell membrane by a glycosyl phosphatidyl inositol anchor, with only small amounts of soluble protein detected. MTF is highly expressed on melanoma cells, and at lower levels in salivary gland, pancreas, kidney and testis. Like other transferrins, MTF is an iron-binding protein, and may play roles in cellular proliferation, tumorigenesis, metastasis and migration. Full-length human, mouse and rat MTF is 738 amino acids (aa). Over aa 1-711, human MTF shares 85 and 86% aa identity with mouse and rat MTF, respectively.
  • Entrez Gene IDs:
    4241 (Human); 30060 (Mouse); 288038 (Rat)
  • Alternate Names:
    antigen p97 (melanoma associated) identified by monoclonal antibodies 133.2 and96.5; CD228 antigen; CD228; FLJ38863; MAP97; MAP97Melanoma-associated antigen p97; Melanotransferrin; membrane-bound transferrin-like protein; MFI2; MGC4856; MTF; MTF1; p97
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