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Human Cystatin B Antibody

Catalog #: MAB1408
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MAB1408
MAB1408-SP

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Detection of Human Cystatin B by Western Blot.
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Human Cystatin B Antibody Summary

Species Reactivity
Human
Specificity
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.
Source
Monoclonal Mouse IgG2B Clone # 225228
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
E. coli-derived recombinant human Cystatin B
aa Met2-Phe98
Accession # P04080
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
2 µg/mL
See below
Immunoprecipitation
25 µg/mL
Conditioned cell culture medium spiked with Recombinant Human Cystatin B (Catalog # 1408-PI), see our available Western blot detection antibodies
Knockout Validated
Cystatin B is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in Cystatin B knockout HEK293T cell line.
 
Neutralization
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.

Scientific Data

Western Blot Detection of Human Cystatin B antibody by Western Blot. View Larger

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 Neutralization of Cystatin B Activity by Human Cystatin B Antibody. View Larger

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.

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

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 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) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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).

References
  1. Abrahamson, M. (1994) Methods Enzymol. 244:685.
  2. Pol, E. and I. Bjork (1999) Biochemistry 38:10519.
  3. Pennacchio, L.A. et al. (1996) Science 271:1731.
  4. Zhang, R. et al. (2003) Glia 42:194.
  5. Zerovnik, E. et al. (2002) Biochim. Biophys. Acta 1594:1.
Entrez Gene IDs
1476 (Human); 13014 (Mouse)
Alternate Names
CPI-B; CST6cystatin B (liver thiol proteinase inhibitor)10STFBcystatin-B; CSTB; cystatin B (stefin B); Cystatin B; EPM1; Liver thiol proteinase inhibitor; PME; Stefin B; Stefin-B

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Citations for Human 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.

3 Citations: Showing 1 - 3
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  1. Insights into the Genetic Profile of Two Siblings Affected by Unverricht-Lundborg Disease Using Patient-Derived hiPSCs
    Authors: Valeria Lucchino, Luana Scaramuzzino, Stefania Scalise, Michela Lo Conte, Clara Zannino, Giorgia Lucia Benedetto et al.
    Cells
  2. Dysregulation of macrophage-secreted cathepsin B contributes to HIV-1-linked neuronal apoptosis.
    Authors: Rodriguez-Franco EJ, Cantres-Rosario YM, Plaud-Valentin M, Romeu R, Rodriguez Y, Skolasky R, Melendez V, Cadilla CL, Melendez LM
    PLoS ONE, 2012-05-31;7(5):e36571.
    Species: Human
    Sample Types: Whole Cells, Whole Tissue
    Applications: ICC, IHC-Fr
  3. Cystatin B as a tissue and urinary biomarker of bladder cancer recurrence and disease progression.
    Authors: Feldman AS, Banyard J, Wu CL, McDougal WS, Zetter BR
    Clin. Cancer Res., 2009-02-01;15(3):1024-31.
    Species: Human
    Sample Types: Urine
    Applications: Western Blot

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