Detection of Human, Mouse, and Rat BLMH/ Bleomycin Hydrolase by Western Blot.
Western blot shows lysates of MOLT‑4 human acute lymphoblastic leukemia cell line, mouse pancreas tissue, and rat pancreas tissue. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human BLMH/Bleomycin Hydrolase Monoclonal Antibody (Catalog # MAB6200) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for BLMH/Bleomycin Hydrolase at approximately 52 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
BLMH/Bleomycin Hydrolase in Human Testis.
BLMH/Bleomycin Hydrolase was detected in immersion fixed paraffin-embedded sections of human testis using Mouse Anti-Human BLMH/Bleomycin Hydrolase Monoclonal Antibody (Catalog # MAB6200) at 15 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 spermatocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 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. 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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