Human Cystatin E/M Antibody

(3 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Cystatin E/M in direct ELISAs and Western blots. In direct ELISAs, approximately 25% cross-reactivity with recombinant mouse Cystatin E/M is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Cystatin E/M
    Arg29-Met149
    Accession # Q15828
  • 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
    1 µg/mL
    See below
  • Simple Western
    50 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    Immersion fixed paraffin-embedded sections of human skin
  • Immunoprecipitation
    25 µg/mL
    Conditioned cell culture medium spiked with Recombinant Human Cystatin E/M (Catalog # 1286-PI), see our available Western blot detection antibodies
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 E/M by Western Blot. Western blot shows lysates of human skin tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Cystatin E/M Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1286) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Cystatin E/M at approximately 13 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Cystatin E/M by Simple WesternTM. Simple Western lane view shows lysates of human skin tissue, loaded at 0.2 mg/mL. A specific band was detected for Cystatin E/M at approximately 13 kDa (as indicated) using 50 µg/mL of Goat Anti-Human Cystatin E/M Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1286) 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. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
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 E/M

Cystatin E/M encoded by the CST6 gene is a member of family 2 of the cystatin superfamily (1, 2). It inhibits papain and cathepsin B, two of the cysteine proteases. Its mRNA was found in many tissues by the two groups who did initial cloning (1, 2). However, its protein was found only in skin and sweat glands by a third group (3). In addition to being a cysteine protease inhibitor, cystatin E/M is also a substrate for transglutaminases (3). It is required for viability and for correct formation of cornified layers in the epidermis and hair follicles, as ichq mice, with a null mutation in the cystatin E/M gene, have defects in epidermal cornification and die between 5 and 12 days of age (4). Cystatin E/M expression and function may not be limited to cutaneous epithelia. For example, it is found in rat brain and is induced during neuronal cell differentiation (5).

  • References:
    1. Sotiropoulou, G. et al. (1997) J. Biol. Chem. 272:903.
    2. Ni, J. et al. (1997) J. Biol. Chem. 272:10853.
    3. Zeeuwen, P.L. et al. (2001) J. Invest. Dermatol. 116:693.
    4. Zeeuwen, P.L. et al. (2002) Hum. Mol. Genet. 11:2867.
    5. Hong, J. et al. (2002) J. Neurochem. 81:922.
  • Entrez Gene IDs:
    1474 (Human); 73720 (Mouse)
  • Alternate Names:
    CST6; cystatin 6; Cystatin E/M; cystatin M; cystatin M/E; Cystatin-6; cystatin-E; cystatin-M; cysteine proteinase inhibitor
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. Differential secretome analysis reveals CST6 as a suppressor of breast cancer bone metastasis.
    Cell Res., 2012;22(9):1356-73.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  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. Invasion suppressor cystatin E/M (CST6): high-level cell type-specific expression in normal brain and epigenetic silencing in gliomas.
    Authors: Qiu J, Ai L, Ramachandran C, Yao B, Gopalakrishnan S, Fields CR, Delmas AL, Dyer LM, Melnick SJ, Yachnis AT, Schwartz PH, Fine HA, Brown KD, Robertson KD
    Lab. Invest., 2008;88(9):910-25.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
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