Cathepsin B Human ELISA Kit (DEIA1416)

Regulatory status: For research use only, not for use in diagnostic procedures.

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Size
96T
Sample
cell culture supernatant, serum, plasma, other biological fluids, tissues extracts
Species Reactivity
Human
Intended Use
For the quantitative measurement of Human Cathepsin B concentrations in serum, plasma body fluids, tissue lysates and cell culture supernatants.
Contents of Kit
1. Recombinant Human Cathepsin B Standard (lyophilized): 2 x 10 ng/tube
2. 96-well plate precoated with anti-human Cathepsin B antibody: 1
3. Sample diluent buffer: 30 mL
4. Biotinylated anti-human Cathepsin B antibody (dilution 1: 100): 130 μL
5. Antibody diluent buffer: 12 mL
6. Avidin-Biotin-Peroxidase Complex (ABC) (dilution 1: 100): 130 μL
7. ABC diluent buffer: 12 mL
8. TMB color developing agent: 10 mL
9. TMB stop solution: 10 mL
Storage
Store the unopened kit at 4°C upon receipt and when it is not in use. For more detailed information, please download the following document on our website.
Detection Range
156 pg/mL-10,000 pg/mL
Sensitivity
5 pg/mL
Standard Curve

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References


Cathepsin proteases promote angiogenic sprouting and laser-induced choroidal neovascularisation in mice

EXPERIMENTAL EYE RESEARCH

Authors: Buehler, A.; Berger, S.; Bengsch, F.; Martin, G.; Han, H.; Vierkotten, S.; Pielen, A.; Boehringer, D.; Schlunck, G.; Fauser, S.; Agostini, H. T.; Reinheckel, T.; Stahl, A.

Cysteine cathepsins are a family of proteases involved in intracellular protein turnover and extracellular matrix degradation. Cathepsin B (Ctsb) and cathepsin Z (Ctsz) promote tumorigenesis and Ctsb is a known modulator of tumor angiogenesis. We therefore investigated the angiomodulatory function of these cathepsins in vitro as well as in a mouse model of laser-induced choroidal neovascularization (laser-CNV). Ctsb(-/-), Ctsz(-/-), Ctsb/Ctsz double-knockout (Ctsb/z DKO), and wild type (WT) mice underwent argon laser treatment to induce choroidal neovascularization (CNV). The neovascularized area was quantified individually for each lesion at 14 days after laser coagulation. In vitro the effects of cathepsin inhibitors on angiogenesis were analysed by endothelial cell (EC) spheroid sprouting and EC invadosome assays. Retinas from cathepsin KO mice did not show gross morphological abnormalities. In the laser CNV model, however, Ctsb/z DKO mice displayed a significantly reduced neovascularized area compared to WT (0.027 mm(2) vs. 0.052 mm(2); p = 0.012), while single knockouts did not differ significantly from WT. In line, VEGF-induced EC spheroid sprouting and invadosome formation were not significantly altered by a specific cathepsin B inhibitor alone, but significantly suppressed when more than one cathepsin was inhibited. Our results demonstrate that laser-CNV formation is significantly reduced in Ctsb/z DKO mice. In line, EC sprouting and invadosome formation are blunted when more than one cathepsin is inhibited in vitro. These results reveal an angiomodulatory potential of cathepsins with partial functional redundancies between different cathepsin family members. (C) 2013 Elsevier Ltd. All rights reserved.

Tumor cell-derived and macrophage-derived cathepsin B promotes progression and lung metastasis of mammary cancer

CANCER RESEARCH

Authors: Vasiljeva, Olga; Papazoglou, Anna; Krueger, Achim; Brodoefel, Harald; Korovin, Matvey; Deussing, Jan; Augustin, Nicole; Nielsen, Boye S.; Almholt, Kasper; Bogyo, Matthew; Peters, Christoph; Reinheckel, Thomas

Proteolysis in close vicinity of tumor cells is a hallmark of cancer invasion and metastasis. We show here that mouse mammary tumor virus-polyoma middle T antigen (PyMT) transgenic mice deficient for the cysteine protease cathepsin B (CTSB) exhibited a significantly delayed onset and reduced growth rate of mammary cancers compared with wild-type PyMT mice. Lung metastasis volumes were significantly reduced in PyMT;ctsb(+/-), an effect that was not further enhanced in PyMT;ctsb(-/-) mice. Furthermore, lung colonization studies of PyMT cells with different CTSB genotypes injected into congenic wild-type mice and in vitro Matrigel invasion assays confirmed a specific role for tumor-derived CTSB in invasion and metastasis. Interestingly, cell surface labeling of cysteine cathepsins by the active site probe DCG-04 detected up-regulation of cathepsin X on PyMT;ctsb(-/-) cells. Treatment of cells with a neutralizing anti-cathepsin X antibody significantly reduced Matrigel invasion of PyMT;ctsb(-/-) cells but did not affect invasion of PyMT;ctsb(+/+) or PyMT;ctsb(+/-) cells, indicating a compensatory function of cathepsin X in CTSB-deficient tumor cells. Finally, an adoptive transfer model, in which ctsb(+/+), ctsb(+/-), and ctsb(-/-) recipient mice were challenged with PYMT;ctsb(+/+) cells, was used to address the role of stroma-derived CTSB in lung metastasis formation. Notably, ctsb(-/-) mice showed reduced number and volume of lung colonies, and infiltrating macrophages showed a strongly up-regulated expression of CTSB within metastatic cell populations. These results indicate that both cancer cell-derived and stroma cell-derived (i.e., macrophages) CTSB plays an important role in tumor progression and metastasis.

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