Detection of Human, Mouse, and Rat PSMA1 by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, Jurkat human acute T cell leukemia cell line, HepG2 human hepatocellular carcinoma cell line, RAW 264.7 mouse monocyte/macrophage cell line, and Rat‑2 rat embryonic fibroblast cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human/Mouse/Rat PSMA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7565) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for PSMA1 at approximately 30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
PSMA1 in Human Breast Cancer Tissue.
PSMA1 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Sheep Anti-Human/Mouse/Rat PSMA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7565) at 1.7 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.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
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.
PSMA-1 (Proteosome subunit alpha type-1; also 30 kDa prosomal protein/PROS30, HC2, proteasome component C2/PSC2, PSMA-1/ alpha 6, and NU) is a 30 kDa member of the peptidase T1A family of enzymes. It is widely expressed, and found in both cytoplasm and nucleus. Short-lived intracellular molecules (typically proteins) are enzymatically degraded by the 26S proteosome. This is a multisubunit 3D complex that is over 2000 kDa in size, and recognizes previously ubiquitinated proteins. The middle of this 26S complex is shaped like a barrel with four staves that run circumferentially rather than longitudinally. Each stave contains seven subunits, with beta ‑type subunits generating the two center staves, and alpha -type subunits comprising the outer, or flanking staves. The function of the barrel, also known as the 20S protease core "particle", is to enzymatically cleave substrates that enter its chamber. For proteins, this is done by beta ‑type subunits. The 26S complex also cleaves mRNA, and this is mediated by alpha -type subunits. PSMA-1/ alpha 6 does not cleave mRNA, but it does positively regulate PSMA5/ alpha 5 catalytic activity. Notably, PSMA-1 has also been reported to bind to LPS. Human PSMA-1 is 263 amino acids (aa) in length. It contains an acetylated Met at position #1, plus three utilized phosphorylation sites at Tyr6, Thr11, and Ser16. There are at least four potential isoform variants. Three utilize alternative start sites. One shows a start site at Met140, while a second and third initiates translation at sites 6 and 43 aa upstream of the standard site, respectively. A fourth isoform possess a 16 aa substitution for aa 115-263. Over aa 16-263, human PSMA-1 shares 98% aa sequence identity with mouse PSMA-1.
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