Detection of Human and Mouse BLMH by Western Blot.
Western blot shows lysates of SH‑SY5Y human neuroblastoma cell line and Neuro‑2A mouse neuroblastoma cell line. PVDF Membrane was probed with 1 µg/mL of Human BLMH Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6200) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for BLMH at approximately 53 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
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.
Background: BLMH/Bleomycin Hydrolase
Bleomycin Hydrolase (BLMH) is a cysteine peptidase of the papain superfamily. It is named for its ability to hydrolyze the antitumor agent bleomycin and inactivate it (1). It has a papain-like catalytic triad (Cys-His-Asp) with optimum activity at neutral pH. In mammals it is expressed ubiquitously in all types of tissues and its expression is up‑regulated in many tumors. It is present in the cytoplasm as homohexameric protein of approximately 300 kDa. In addition to its aminopeptidase activity, it has homocysteine-thiolactonase activity. The normal physiological function of BLMH is not clear. BLMH inactivates bleomycin, a glycopeptide anticancer agent, by deaminating it (2). BLMH has been suggested to play a role in the generation of MHC class I-presented peptides (3 ‑ 4). Diminished BLMH activity may contribute to the pathology of Alzheimer’s disease (AD) (5 ‑ 6). It is inhibited by cysteine protease inhibitors such as N-ethylmaleimide, iodoacetamide, para-hydroxymercuribenzoate, and E-64.
Joshua-Tor, L. and S. A. Johnson (2004) in Handbook of Proteolytic Enzymes, Barrett, A. J. et al. eds. pp. 1197.
Schwartz, D. R. et al. (1999) Proc. Natl. Acad. Sci. USA, 96:4680.
Kim, E. et al. (2009) J. Immunol. 183:7379.
Towne, C. F. et al. (2007) J. Immunol. 178:6923.
Suszynska, J. et al. (2010) J. Alzheimers Dis. 19:1177.
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