Human Cystatin C Antibody

(3 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Cystatin C in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 50% cross-reactivity with recombinant mouse Cystatin C is observed and less than 5% cross-reactivity with recombinant human (rh) Fetuin A, rhFetuin B, rhCystatin A, rhCystatin B, rhCystatin S, and rhCystatin E/M is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Cystatin C
    Ser27-Ala146
    Accession # P01034
  • Formulation
    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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.25-2 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunoprecipitation
    25 µg/mL
    Conditioned cell culture medium spiked with Recombinant Human Cystatin C (Catalog # 1196-PI), see our available Western blot detection antibodies
  • Neutralization
    Measured by its ability to neutralize Recombinant Human Cystatin C (0.64 µg/mL, Catalog # 1196-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 7.8 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.
Data Examples

Detection of Human Cystatin C by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, human liver tissue, and human serum. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Human Cystatin C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1196) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Cystatin C at approximately 14 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Cystatin C by Western Blot. Western blot shows lysates of human placenta tissue and PC‑3 human prostate cancer cell line. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human Cystatin C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1196) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Cystatin C at approximately 14 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry

Cystatin C in Human Breast. Cystatin C was detected in immersion fixed paraffin-embedded sections of human breast using 5 µg/mL Goat Anti-Human Cystatin C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1196) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the cytoplasm in cells of terminal ductules. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    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 C

Cystatin C is a member of family 2 of the Cystatin superfamily (1). It is involved in processes such as tumor invasion and metastasis, inflammation and some neurological diseases. It inhibits many cysteine proteases such as papain and cathepsins B, H, K, L, and S (2, 3). It is ubiquitous in human tissues and body fluids. A point mutation in the gene coding for the 120 amino acid mature Cystatin C causes a hereditary form of amyloid angiopathy in which the protein variant (Leu68 to Gln) is deposited in the cerebral arteries, leading to fatal cerebral hemorrhage (4). Cystatin C may have additional clinical applications. For example, it is a good marker for glomerular filtration rate (5).

  • References:
    1. Reed, C.H. (2000) British J. Biomed. Sci. 57:323.
    2. Janowski, R. et al. (2001) Nat. Struct. Biol. 8:316.
    3. Abrahamson, M. (1994) Methods Enzymol. 244:685.
    4. Abrahamson, M. et al. (1992) Hum. Genet. 89:377.
    5. Laterza, O.F. et al. (2002) Clin. Chem. 48:699.
  • Entrez Gene IDs:
    1471 (Human); 13010 (Mouse); 25307 (Rat)
  • Alternate Names:
    ARMD11; bA218C14.4 (cystatin C); CST3; cystatin 3; cystatin C (amyloid angiopathy and cerebral hemorrhage); Cystatin C; cystatin-3; cystatin-C; Gamma-trace; MGC117328; Neuroendocrine basic polypeptide; Post-gamma-globulin
Related Research Areas
Citations:

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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Species
Applications
Sample Type
  1. Nanoparticle Detection of Urinary Markers for Point-of-Care Diagnosis of Kidney Injury.
    Authors: Chung H, Pellegrini K, Chung J, Wanigasuriya K, Jayawardene I, Lee K, Lee H, Vaidya V, Weissleder R
    PLoS ONE, 2015;10(7):e0133417.
    Species: Human
    Sample Type: Urine
    Application: ELISA Development
  2. Cystatin E/M suppresses legumain activity and invasion of human melanoma.
    Authors: Briggs JJ, Haugen MH, Johansen HT, Riker AI, Abrahamson M, Fodstad O, Maelandsmo GM, Solberg R
    BMC Cancer, 2010;10(1):17.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. Proteomic analysis of factors released from p21-overexpressing tumour cells.
    Authors: Currid CA, O'Connor DP, Chang BD, Gebus C, Harris N, Dawson KA, Dunn MJ, Pennington SR, Roninson IB, Gallagher WM
    Proteomics, 2006;6(13):3739-53.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: Array Development
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