Human Metadherin Antibody

Catalog # Availability Size / Price Qty
AF7644
AF7644-SP
Detection of Human Metadherin by Western Blot.
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Human Metadherin Antibody Summary

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
Immunohistochemistry
3-25 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

Western Blot Detection of Human Metadherin by Western Blot. View Larger

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. View Larger

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.

Immunohistochemistry Metadherin  in Human Breast. View Larger

Metadherin in Human Breast. Metadherin was detected in immersion fixed paraffin-embedded sections of human breast using Sheep Anti-Human Metadherin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7644) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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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FAQs

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Cell and Tissue Staining Kits

Immunohistochemistry Reagents

Isotype Controls

Reconstitution Buffers

Secondary Antibodies

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