Human BLMH/Bleomycin Hydrolase Antibody

Catalog # Availability Size / Price Qty
MAB6200
MAB6200-SP
Detection of Human, Mouse, and Rat BLMH/ Bleomycin Hydrolase by Western Blot.
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Product Details
Citations (1)
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Human BLMH/Bleomycin Hydrolase Antibody Summary

Species Reactivity
Human
Specificity
Detects human BLMH/Bleomycin Hydrolase in direct ELISAs. It detects human, mouse and rat BLMH/Bleomycin Hydrolase in Western Blots.
Source
Monoclonal Mouse IgG2B Clone # 760239
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
E. coli-derived recombinant human BLMH/Bleomycin Hydrolase
Ser2-Glu455
Accession # Q13867
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.2 µg/mL
See below
Immunohistochemistry
8-25 µg/mL
See below
Knockout Validated
BLMH/Bleomycin Hydrolase is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in BLMH/Bleomycin Hydrolase knockout HeLa cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human, Mouse, and Rat BLMH/ Bleomycin Hydrolase antibody by Western Blot. View Larger

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.

Immunohistochemistry BLMH/Bleomycin Hydrolase antibody in Human Testis by Immunohistochemistry (IHC-P). View Larger

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.

Knockout Validated Western Blot Shows Human BLMH/Bleomycin Hydrolase Antibody Specificity by Using Knockout Cell Line. View Larger

Western Blot Shows Human BLMH/Bleomycin Hydrolase Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and BLMH/Bleomycin Hydrolase knockout HeLa cell line (KO). 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 51 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.5 mg/mL.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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.

References
  1. Joshua-Tor, L. and S. A. Johnson (2004) in Handbook of Proteolytic Enzymes, Barrett, A. J. et al. eds. pp. 1197.
  2. Schwartz, D. R. et al. (1999) Proc. Natl. Acad. Sci. USA, 96:4680.
  3. Kim, E. et al. (2009) J. Immunol. 183:7379.
  4. Towne, C. F. et al. (2007) J. Immunol. 178:6923.
  5. Suszynska, J. et al. (2010) J. Alzheimers Dis. 19:1177.
  6. Lefterov, I. M. et al. (2000) FASEB J. 14:1837.
Entrez Gene IDs
642 (Human)
Alternate Names
BHBMHBLM hydrolase; bleomycin hydrolase; BLMH; BMH; EC 3.4.22.40

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Citation for Human BLMH/Bleomycin Hydrolase Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Nrf2 mediates the resistance of human A549 and HepG2 cancer cells to boningmycin, a new antitumor antibiotic, in vitro through regulation of glutathione levels
    Authors: HX Zhang, Y Chen, R Xu, QY He
    Acta Pharmacol. Sin., 2018-05-10;39(10):1661-1669.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot

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