hh Human Cathepsin B Antibody AF953: R&D Systems

Human Cathepsin B Antibody

(13 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Cathepsin B in direct ELISAs and Western blots. In direct ELISAs, approximately 35% cross-reactivity with recombinant mouse (rm) Cathepsin B is observed and less than 5% cross-reactivity with recombinant human (rh) Cathepsin C, rmCathepsin H, and rhCathepsin L is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Cathepsin B
    Arg18-Ile339
    Accession # P07858
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.25 µg/mL
    See below
  • Simple Western
    2.5 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunoprecipitation
    25 µg/mL
    Conditioned cell culture medium spiked with Recombinant Human Cathepsin B (Catalog # 953-CY), see our available Western blot detection antibodies
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.
Data Examples
Detection of Human Cathepsin B by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Human Cathepsin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF953) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Cathepsin B at approximately 25-30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunohistochemistry
Cathepsin B in Human Brain. Cathepsin B was detected in immersion fixed paraffin-embedded sections of human brain (cortex) using Goat Anti-Human Cathepsin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF953) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human Cathepsin B by Simple WesternTM. Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Cathepsin B at approximately 34 kDa (as indicated) using 2.5 µg/mL of Goat Anti-Human Cathepsin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF953) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    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: Cathepsin B

Cathepsin B is the first described member of the family of lysosomal cysteine proteases (1). Cathepsin B possesses both endopeptidase and exopeptidase activities, in the latter case acting as a peptidyl-dipeptidase. It is known to process a number of proteins, including pro and active caspases, prorenin, and secretory leucoprotease inhibitor (SLPI) (2-4). Therefore, Cathepsin B may play a role in activation and inactivation of caspases, activation of renin and inactivation of SLPI, the key steps in apoptosis, angiotensin production, and progression of emphysema, respectively. Because of its increased levels and redistribution of the enzyme in human and animal tumors, Cathepsin B may also have role in invasion and metastasis (5).

In addition to lysosome, Cathepsin B can be secreted or associated with plasma membrane, cytoplasm, and nucleus. It is synthesized as a preproenzyme. Following removal of the signal peptide, the inactive proenzyme undergoes further modifications including removal of the pro region to result in the active enzyme (1).

  • References:
    1. Mort, J.S. (2004) in Handbook of Proteolytic Enzymes. Barrett, A.J. et al. (eds): Academic Press, San Diego, p. 1079.
    2. Vancompernolle, K. et al. (1998) FEBS Lett. 438:150.
    3. Jutras, I. and T.L. Reudelhuber (1999) FEBS Lett. 443:48.
    4. Taggart, C.C. et al. (2001) J. Biol. Chem. 276:33345.
    5. Bergquin, I.M. and B.F. Sloane (1996) Adv. Exp. Med. Biol. 389:281.
  • Entrez Gene IDs:
    1508 (Human); 13030 (Mouse)
  • Alternate Names:
    APP secretase; APPS; Cathepsin B; Cathepsin B1; CPSBamyloid precursor protein secretase; CTSB; cysteine protease; EC 3.4.22; EC 3.4.22.1
Related Research Areas
Citations:

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.

13 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. The lysosomal protein cathepsin L is a progranulin protease
    Authors: CW Lee, JN Stankowski, J Chew, CN Cook, YW Lam, S Almeida, Y Carlomagno, KF Lau, M Prudencio, FB Gao, M Bogyo, DW Dickson, L Petrucelli
    Mol Neurodegener, 2017;12(1):55.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. The Unusual Resistance of Avian Defensin AvBD7 to Proteolytic Enzymes Preserves Its Antibacterial Activity
    PLoS ONE, 2016;11(8):e0161573.
    Species: Chicken
    Sample Type: Tissue Homogenates
    Application: WB
  3. Cathepsin S attenuates endosomal EGFR signalling: A mechanical rationale for the combination of cathepsin S and EGFR tyrosine kinase inhibitors
    Sci Rep, 2016;6(0):29256.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  4. Simvastatin inhibits glucose metabolism and legumain activity in human myotubes.
    Authors: Smith R, Solberg R, Jacobsen L, Voreland A, Rustan A, Thoresen G, Johansen H
    PLoS ONE, 2014;9(1):e85721.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  5. Human bone marrow-derived mesenchymal stem cells suppress human glioma growth through inhibition of angiogenesis.
    Authors: Ho I, Toh H, Ng W, Teo Y, Guo C, Hui K, Lam P
    Stem Cells, 2013;31(1):146-55.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Frozen
  6. Differential secretome analysis reveals CST6 as a suppressor of breast cancer bone metastasis.
    Cell Res., 2012;22(9):1356-73.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  7. A possible contribution of altered cathepsin B expression to the development of skin sclerosis and vasculopathy in systemic sclerosis.
    Authors: Noda S, Asano Y, Akamata K, Aozasa N, Taniguchi T, Takahashi T, Ichimura Y, Toyama T, Sumida H, Yanaba K, Tada Y, Sugaya M, Kadono T, Sato S
    PLoS ONE, 2012;7(2):e32272.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  8. Transgenic expression of human cathepsin B promotes progression and metastasis of polyoma-middle-T-induced breast cancer in mice.
    Authors: Sevenich L, Werner F, Gajda M, Schurigt U, Sieber C, Muller S, Follo M, Peters C, Reinheckel T
    Oncogene, 2011;30(1):54-64.
    Species: Human
    Sample Type: Tissue Homogenates
    Application: WB
  9. CD40 stimulation sensitizes CLL cells to lysosomal cell death induction by type II anti-CD20 mAb GA101.
    Authors: Jak M, van Bochove GG, Reits EA
    Blood, 2011;118(19):5178-88.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  10. Macrophages and cathepsin proteases blunt chemotherapeutic response in breast cancer.
    Authors: Shree T, Olson OC, Elie BT
    Genes Dev., 2011;25(23):2465-79.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  11. The NALP3 inflammasome is involved in the innate immune response to amyloid-beta.
    Authors: Halle A, Hornung V, Petzold GC, Stewart CR, Monks BG, Reinheckel T, Fitzgerald KA, Latz E, Moore KJ, Golenbock DT
    Nat. Immunol., 2008;9(8):857-65.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC
  12. Distinct roles for cysteine cathepsin genes in multistage tumorigenesis.
    Authors: Gocheva V, Zeng W, Ke D, Klimstra D, Reinheckel T, Peters C, Hanahan D, Joyce JA
    Genes Dev., 2006;20(5):543-56.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  13. Biomarker discovery from pancreatic cancer secretome using a differential proteomic approach.
    Authors: Gronborg M, Kristiansen TZ, Iwahori A, Chang R, Reddy R, Sato N, Molina H, Jensen ON, Hruban RH, Goggins MG, Maitra A, Pandey A
    Mol. Cell Proteomics, 2006;5(1):157-71.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
Expand to show all 13 Citations
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