Detects human Cystatin B in Western blots and direct ELISAs. In Western blots, less than 5% cross-reactivity with recombinant human (rh) Cystatins A and S, recombinant mouse Cystatin B, rhFetuin A, and rhHPRG (Histidine-Proline-Rich Glycoprotein) is observed and approximately 5% to 10% cross-reactivity with rhCystatins C, D, E/M, SA, SN, and rhFetuin B is observed.
Monoclonal Mouse IgG2B Clone # 225228
Protein A or G purified from hybridoma culture supernatant
E. coli-derived recombinant human Cystatin B aa Met2-Phe98 Accession # P04080
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.
Measured by its ability to neutralize Recombinant Human Cystatin B (0.53 µg/mL, Catalog # 1408-PI) inhibition of Papain (0.1 µg/mL) cleavage of the fluorogenic peptide substrate Z-FR-AMC (100 µM, Catalog # ES009). The Neutralization Dose (ND50) is typically 5.4 µg/mL.
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 Cystatin B by Western Blot. Western blot shows lysates of human colon tissue, SK‑BR‑3 human breast cancer cell line, and A549 human lung carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Cystatin B Monoclonal Antibody (Catalog # MAB1408) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Cystatin B at approximately 12 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Neutralization of Cystatin B Activity by Human Cystatin B Antibody. Papain (0.1 µg/mL) activity is measured in the presence of Recombinant Human Cystatin B (0.53 µg/mL, Catalog # 1408-PI) that has been preincubated with increasing concentrations of Human Cystatin B Monoclonal Antibody (Catalog # MAB1408). The ND50 is typically 5.4 µg/mL.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
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: 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.
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.
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