Detects human Cystatin E/M in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross‑reactivity with recombinant human (rh) Cystatins A, B, C, S, SA, SN, rhFetuin A, B, rhHRGP, or rhKininogen is observed.
Monoclonal Mouse IgG2B Clone # 211515
Protein A or G purified from hybridoma culture supernatant
Mouse myeloma cell line NS0-derived recombinant human Cystatin E/M Arg29-Met149 Accession # Q15828
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 E/M (0.32 µg/mL, Catalog # 1286-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 µg/mL.
Please Note: Optimal dilutions should be determined by each laboratory for each application.
are available in the Technical Information section on our website.
Detection of Human Cystatin E/M by Western Blot.
Western blot shows lysates of human skin tissue and human colon cancer tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Cystatin E/M Monoclonal Antibody (Catalog # MAB1286) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). 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.
Cystatin E/M was detected in immersion fixed paraffin-embedded sections of human skin using 25 µg/mL Mouse Anti-Human Cystatin E/M Monoclonal Antibody (Catalog # MAB1286) overnight at 4 °C. Tissue was stained with the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific labeling was localized to the cytoplasm of cells in hair follicules. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
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).
Sotiropoulou, G. et al. (1997) J. Biol. Chem. 272:903.
Ni, J. et al. (1997) J. Biol. Chem. 272:10853.
Zeeuwen, P.L. et al. (2001) J. Invest. Dermatol. 116:693.
Zeeuwen, P.L. et al. (2002) Hum. Mol. Genet. 11:2867.
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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