Human Cathepsin B Antibody

Catalog # Availability Size / Price Qty
AF953
AF953-SP
Detection of Human Cathepsin B by Western Blot.
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Product Details
Citations (18)
FAQs
Supplemental Products
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.

Data Examples

Western Blot Detection of Human Cathepsin B 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 in Human Brain. 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 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 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 Detection of Cathepsin B in U‑87 (Positive) & Daudi (Negative). 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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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.

18 Citations: Showing 1 - 10
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  1. 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;0(0):.
    Species: Gerbil
    Sample Types: Cell Lysates
    Applications: Western Blot
  2. 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;10(2):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. 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;12(9):.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  4. 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;0(0):1-14.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  5. 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;38(23):5399-5414.
    Species: Rat
    Sample Types: Cell Culture Supernates
    Applications: Western Blot
  6. 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 Types: Cell Lysates
    Applications: Western Blot
  7. The Unusual Resistance of Avian Defensin AvBD7 to Proteolytic Enzymes Preserves Its Antibacterial Activity
    PLoS ONE, 2016;11(8):e0161573.
    Species: Chicken
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  8. 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 Types: Cell Lysates
    Applications: Western Blot
  9. 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 Types: Cell Lysates
    Applications: Western Blot
  10. 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 Types: Whole Tissue
    Applications: IHC-Fr
  11. Differential secretome analysis reveals CST6 as a suppressor of breast cancer bone metastasis.
    Cell Res., 2012;22(9):1356-73.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  12. 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 Types: Whole Tissue
    Applications: IHC-P
  13. 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 Types: Cell Lysates
    Applications: Western Blot
  14. 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 Types: Tissue Homogenates
    Applications: Western Blot
  15. 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 Types: Whole Tissue
    Applications: IHC-P
  16. 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 Types: Whole Cells
    Applications: ICC
  17. 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 Types: Cell Lysates
    Applications: Western Blot
  18. 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 Types: Whole Tissue
    Applications: IHC-P

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Human Cathepsin B Antibody
By Anonymous on 08/06/2021
Application: 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