Detection of Mouse Metadherin by Western Blot. Western blot shows lysates of 4T1 mouse breast cancer cell line and P19 mouse embryonal carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Rat Anti-Mouse Metadherin Monoclonal Antibody (Catalog # MAB7180) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Metadherin at approximately 80 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
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.
Metadherin (MTDH; also Lyric and Astrocyte-elevated gene-1/AEG1) is a unique molecule originally discovered to be upregulated in astrocytes following HIV infection. Although its predicted MW is 64 kDa, it runs anomalously at approximately 80 kDa in SDS-Page. It is widely expressed, and appears to be a component of the ER, nucleolus and inner nuclear membrane. Metadherin is reported to promote Akt/PI3 kinase activity, and suppress apoptosis-associated FOXO3A transcription. Mouse Metadherin is a type III (i.e.- a type I with no signal sequence) transmembrane protein 579 amino acids (aa) in length. It contains a luminal N-terminus (aa 1-48) with a 510 aa cytoplasmic C-terminus. There are no identifiable structural motifs, although two poly-Lys segments and two utilized Ser phosphorylation sites exist. Multiple bands at 80 kDa, 75 kDa, 50-55 kDa and 37 kDa are seen for Metadherin on SDS-Page. They may reflect the presence of at least two potential isoform variants that show 1) a deletion of aa 422-457, and 2) an Ile substitution for aa 190-579. Over aa 167-297, mouse Metadherin shares 98% and 97% aa sequence identity with rat and human Metadherin, respectively.
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