Detection of Human Kallikrein 3/PSA by Western Blot.
Western blot shows lysates of human prostate tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Kallikrein 3/PSA Monoclonal Antibody (Catalog # MAB1344) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Kallikrein 3/PSA at approximately 30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Kallikrein 3/PSA in LNCaP and MCF-7 Human Cell Lines.
Kallikrein 3/PSA was detected in immersion fixed LNCaP human prostate cancer cell line (positive control, left panel) and MCF‑7 human breast cancer cell line (negative control, right panel) using Mouse Anti-Human Kallikrein 3/PSA Monoclonal Antibody (Catalog # MAB1344) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm in LNCaP cells. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Kallikrein 3/PSA in Prostate Cancer.
Kallikrein 3/PSA was detected in immersion fixed paraffin-embedded sections of prostate cancer tissue using Mouse Anti-Human Kallikrein 3/PSA Monoclonal Antibody (Catalog # MAB1344) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to the apical plasma membrane. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
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.
Background: Kallikrein 3/PSA
Kallikrein 3, commonly known as prostate specific antigen (PSA), is a serine protease of the human tissue Kallikrein gene family (1). PSA is synthesized in the ductal and acinar epithelium of the prostate gland and secreted into the seminal plasma in high concentrations (0.5-2 g/L) (2). A small portion of PSA “leaks” into the systemic circulation, the levels of which increase significantly (30-fold) from prostate cancer tissue than normal prostate tissue (3). PSA has become a well established tumor marker that aids the diagnosis, staging, and follow up of prostate cancer. The deduced amino acid sequence of human PSA consists of a signal peptide, a short pro region and a mature/active enzyme. The pro-enzyme is activated, possibly by active Kallikreins 2, 4 or 15 in vivo (4). Recombinant human PSA is activated by thermolysin, a zinc protease. The active PSA cleaves several tyrosyl peptide bonds in semenogelins I and II, which are the major gel-forming proteins produced by the seminal vesicles (5). Several inhibitors including serpin A3/ alpha 1-antichymotrypsin (ACT) and alpha 2-macroglobulin are known to form complexes with PSA.
Yousef, G.M. and E.P. Diamandis (2001) Endocrine Rev. 22:184.
Ward, A.M. et al. (2001) Ann. Clin. Biochem. 38:633.
Jain, S. et al. (2002) Postgrad. Med. J. 78:646.
Lilja H. (2003) Urology 62:270.
Takayama, T.K. et al. (1997) J. Biol. Chem. 272:21582.
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