Human Cathepsin B Antibody

Catalog # Availability Size / Price Qty
AF953
AF953-SP

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Detection of Human Cathepsin B by Western Blot.
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Product Details
Citations (29)
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Reviews (3)

Human Cathepsin B Antibody Summary

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 either lyophilized or 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
Immunocytochemistry
5-25 µg/mL
Immersion fixed U‑87 MG human glioblastoma/astrocytoma cell line (Positive) & Daudi human Burkitt's lymphoma cell line (Negative)
Knockout Validated
Cathepsin B is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in Cathepsin B 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 Cathepsin B antibody by Western Blot. View Larger

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 antibody in Human Brain by Immunohistochemistry (IHC-P). View Larger

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; 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.

Simple Western Detection of Human Cathepsin B antibody by Simple Western<SUP>TM</SUP>. View Larger

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 (HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Knockout Validated Western Blot Shows Human Cathepsin B Antibody Specificity by Using Knockout Cell Line. View Larger

Western Blot Shows Human Cathepsin B Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Cathepsin B knockout HeLa cell line (KO). 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 (HAF017). Specific bands were detected for Cathepsin B at approximately 25-45 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1. New adjunct appears with knockout cell line.

Immunocytochemistry View Larger

Detection of Cathepsin B in U‑87 (Positive) & Daudi (Negative). Cathepsin B was detected in immersion fixed U‑87 MG human glioblastoma/astrocytoma cell line (Positive) & absent in Daudi human Burkitt's lymphoma cell line (Negative) using Goat Anti-Human Cathepsin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF953) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cell surface. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Western Blot Detection of Human Cathepsin B by Western Blot View Larger

Detection of Human Cathepsin B by Western Blot Legumain, cathepsin B and L expressions in cytosolic and membrane fractions of myotubes.Myotubes were treated with (+) or without (−) 30 µM simvastatin for 48 h before subcellular fractionation was performed. One representative immunoblot of legumain, cathepsin B and L in the cytosolic and membrane fractions is shown. All lanes were loaded with equal amount of total proteins and probed with antibodies as indicated. LAMP-2 and alpha -tubulin are shown as cell compartment controls (n = 3). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24416446), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Human Cathepsin B by Western Blot View Larger

Detection of Human Cathepsin B by Western Blot CTSS inhibition does not impair lysosomal activities.(a) OEC-M1 cells were pretreated with vesicle or 100 nM BAF for 1 h and then incubated with 20 μM 6r for 1 h further. Lysosomal proteolytic activities were determined using BODIPY–BSA and quantified through flow cytometry. (b) After 48 h of siRNA knockdown of CTSS and CTSB, the relative expression of CTSS and CTSB were determined through Western blotting (right panel). Lysosomal proteolytic activities were determined using BODIPY–BSA and quantified through flow cytometry (right panel). Data represent the mean ± SD of three independent experiments. Differences were found to be statistically significant at ***P < 0.001. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/srep29256), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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

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Citations for Human Cathepsin B 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.

29 Citations: Showing 1 - 10
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  1. Reciprocal effects of alpha-synuclein aggregation and lysosomal homeostasis in synucleinopathy models
    Authors: Drobny A, Boros FA, Balta D et al.
    Translational neurodegeneration
  2. Salivary gland LAMP3 mRNA expression is a possible predictive marker in the response to hydroxychloroquine in Sj�gren's disease
    Authors: H Nakamura, T Tanaka, Y Ji, C Zheng, SA Afione, BM Warner, FR Oliveira, ACF Motta, EM Rocha, M Noguchi, T Atsumi, JA Chiorini
    PLoS ONE, 2023-02-23;18(2):e0282227.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. Autolysosomal activation combined with lysosomal destabilization efficiently targets myeloid leukemia cells for cell death
    Authors: Harshit Shah, Metodi Stankov, Diana Panayotova-Dimitrova, Amir Yazdi, Ramachandramouli Budida, Jan-Henning Klusmann et al.
    Frontiers in Oncology
  4. Current methods to analyze lysosome morphology, positioning, motility and function
    Authors: Duarte C. Barral, Leopoldo Staiano, Cláudia Guimas Almeida, Dan F. Cutler, Emily R. Eden, Clare E. Futter et al.
    Traffic
  5. An infection-induced oxidation site regulates legumain processing and tumor growth
    Authors: Y Kovalyova, DW Bak, EM Gordon, C Fung, JHB Shuman, TL Cover, MR Amieva, E Weerapana, SK Hatzios
    Nature Chemical Biology, 2022-03-24;0(0):.
    Species: Gerbil
    Sample Types: Cell Lysates
    Applications: Western Blot
  6. ORF3a of SARS-CoV-2 promotes lysosomal exocytosis-mediated viral egress
    Authors: Di Chen, Qiaoxia Zheng, Long Sun, Mingming Ji, Yan Li, Hongyu Deng et al.
    Developmental Cell
  7. NDST3 deacetylates alpha ‐tubulin and suppresses V‐ATPase assembly and lysosomal acidification
    Authors: Qing Tang, Mingming Liu, Yang Liu, Ran‐Der Hwang, Tao Zhang, Jiou Wang
    The EMBO Journal
  8. Impact of DJ-1 and Helix 8 on the Proteome and Degradome of Neuron-Like Cells
    Authors: U Kern, K Fröhlich, J Bedacht, N Schmidt, ML Biniossek, N Gensch, K Baerenfall, O Schilling
    Cells, 2021-02-16;10(2):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  9. ORF3a of the COVID-19 virus SARS-CoV-2 blocks HOPS complex-mediated assembly of the SNARE complex required for autolysosome formation
    Authors: Guangyan Miao, Hongyu Zhao, Yan Li, Mingming Ji, Yong Chen, Yi Shi et al.
    Developmental Cell
  10. The secreted inhibitor of invasive cell growth CREG1 is negatively regulated by cathepsin proteases
    Authors: Alejandro Gomez-Auli, Larissa Elisabeth Hillebrand, Daniel Christen, Sira Carolin Günther, Martin Lothar Biniossek, Christoph Peters et al.
    Cellular and Molecular Life Sciences
  11. Cathepsin Inhibition Modulates Metabolism and Polarization of Tumor-Associated Macrophages
    Authors: D Oelschlaeg, T Weiss Sada, S Salpeter, S Krug, G Blum, W Schmitz, A Schulze, P Michl
    Cancers (Basel), 2020-09-10;12(9):.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  12. Myocardial cathepsin D is downregulated in sudden cardiac death
    Authors: Yu Kakimoto, Ayumi Sasaki, Maki Niioka, Noboru Kawabe, Motoki Osawa
    PLOS ONE
  13. (-)-Oleocanthal and (-)-oleocanthal-rich olive oils induce lysosomal membrane permeabilization in cancer cells
    Authors: Limor Goren, George Zhang, Susmita Kaushik, Paul A. S. Breslin, Yi-Chieh Nancy Du, David A. Foster
    PLOS ONE
  14. YWHA/14-3-3 proteins recognize phosphorylated TFEB by a noncanonical mode for controlling TFEB cytoplasmic localization
    Authors: Y Xu, J Ren, X He, H Chen, T Wei, W Feng
    Autophagy, 2019-01-27;0(0):1-14.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  15. Modulation of Receptor Protein Tyrosine Phosphatase Sigma Increases Chondroitin Sulfate Proteoglycan Degradation through Cathepsin B Secretion to Enhance Axon Outgrowth
    Authors: AP Tran, S Sundar, M Yu, BT Lang, J Silver
    J. Neurosci., 2018-05-14;38(23):5399-5414.
    Species: Rat
    Sample Types: Cell Culture Supernates, Whole Cells, Whole Tissue
    Applications: ICC, IHC, Western Blot
  16. Abl and Arg mediate cysteine cathepsin secretion to facilitate melanoma invasion and metastasis
    Authors: Rakshamani Tripathi, Leann S. Fiore, Dana L. Richards, Yuchen Yang, Jinpeng Liu, Chi Wang et al.
    Science Signaling
  17. 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, 2017-07-25;12(1):55.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  18. The Unusual Resistance of Avian Defensin AvBD7 to Proteolytic Enzymes Preserves Its Antibacterial Activity
    PLoS ONE, 2016-08-25;11(8):e0161573.
    Species: Chicken
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  19. Cathepsin S attenuates endosomal EGFR signalling: A mechanical rationale for the combination of cathepsin S and EGFR tyrosine kinase inhibitors
    Sci Rep, 2016-07-08;6(0):29256.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  20. 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, 2014-01-08;9(1):e85721.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: ICC, Western Blot
  21. 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, 2013-01-01;31(1):146-55.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-Fr
  22. Differential secretome analysis reveals CST6 as a suppressor of breast cancer bone metastasis.
    Cell Res., 2012-06-12;22(9):1356-73.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  23. 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, 2012-02-23;7(2):e32272.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  24. Macrophages and cathepsin proteases blunt chemotherapeutic response in breast cancer.
    Authors: Shree T, Olson OC, Elie BT
    Genes Dev., 2011-12-01;25(23):2465-79.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  25. 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, 2011-09-26;118(19):5178-88.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  26. 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, 2010-09-06;30(1):54-64.
    Species: Human
    Sample Types: Tissue Homogenates, Whole Tissue
    Applications: IHC-P, Western Blot
  27. 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., 2008-07-11;9(8):857-65.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: ICC
  28. 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., 2006-02-15;20(5):543-56.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  29. 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, 2005-10-08;5(1):157-71.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot

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Reviews for Human Cathepsin B Antibody

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Human Cathepsin B Antibody
By Anonymous on 08/06/2021
Application: WB
Reason for Rating: This antibody works very good in human kidney cell lines Sample Tested: renal carcinoma Species: Human

1:500 dilution is good.


Human Cathepsin B Antibody
By Anonymous on 08/06/2021
Application: It worked very good in human kidney cell lines Sample Tested: renal cancer cell,Renal cancer tissue Species: Human

It's better to use 12% SDS-PAGE


Human Cathepsin B Antibody
By Anonymous on 11/12/2020
Application: WB Sample Tested: ARP-1 human myeloma cell line Species: Human