Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Knockout Validated, Western Blot, Neutralization, Immunoprecipitation

Cited:

Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 225228
View all Cystatin B Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Cystatin B
aa Met2-Phe98
Accession # P04080

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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human Cystatin B Antibody (225228)

Detection of Human Cystatin B antibody by Western Blot.

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.

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

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.

Applications for Human Cystatin B Antibody (225228)

Application
Recommended Usage

Immunoprecipitation

25 µg/mL
Sample: 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.

Western Blot

2 µg/mL
Sample:   Human colon tissue, SK‑BR‑3 human breast cancer cell line, and A549 human lung carcinoma 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.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

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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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.

Alternate Names

CSTB, Stefin B

Entrez Gene IDs

1476 (Human); 13014 (Mouse)

Gene Symbol

CSTB

UniProt

P04080

Additional Cystatin B Products

Product Documents for Human Cystatin B Antibody (225228)

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human Cystatin B Antibody (225228)

For research use only

Related Research Areas

Cysteine Proteases and Regulators

Citations for Human Cystatin B Antibody (225228)

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