Human/Rat Cystatin B Antibody Summary
Accession # P04080
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.
Detection of Human and Rat Cystatin B by Western Blot. Western blot shows lysates of human colon tissue, PC‑12 rat adrenal pheochromocytoma cell line, A549 human lung carcinoma cell line, and SK‑BR‑3 human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Rat Cystatin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1408) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Cystatin B at approximately 11 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Cystatin B by Simple WesternTM.
Simple Western lane view shows lysates of A549 human lung carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Cystatin B at approximately 18 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Rat Cystatin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1408) 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.
Western Blot Shows Human Cystatin B Specificity by Using Knockout Cell Line. Western blot shows lysates of HEK293T human embryonic kidney parental cell line and Cystatin B knockout HEK293T cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Rat Cystatin B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1408) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Cystatin B at approximately 12 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
- 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: Cystatin B
Cystatin B, also called stefin B or liver thiol proteinase inhibitor, is a member of family 1 of the cystatin superfamily (1). Like Cystatin A, it is an intracellular inhibitor regulating the activities of cysteine proteases of the papain family such as cathepsins B, H and L (2). Mutations in the Cystatin B gene is the cause of progressive myoclonus epilepsy (EPM1) (3). Because of its expression patterns, Cystatin B can be used as a marker for certain cancers, such as glioblastoma tumors (4). It readily forms amyloid fibrils in vitro (5). The human Cystatin B consists of 98 amino acid residues (3).
- Abrahamson, M. (1994) Methods Enzymol. 244:685.
- Pol, E. and I. Bjork (1999) Biochemistry 38:10519.
- Pennacchio, L.A. et al. (1996) Science 271:1731.
- Zhang, R. et al. (2003) Glia 42:194.
- Zerovnik, E. et al. (2002) Biochim. Biophys. Acta 1594:1.
Citation for Human/Rat Cystatin 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.
1 Citation: Showing 1 - 1
Regulation of TGF-beta1-driven differentiation of human lung fibroblasts: emerging roles of cathepsin B and cystatin C.
Authors: Kasabova M, Joulin-Giet A, Lecaille F, Gilmore B, Marchand-Adam S, Saidi A, Lalmanach G
J Biol Chem, 2014;289(23):16239-51.
Sample Types: Cell Lysates
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