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Human Cystatin E/M Antibody

Catalog #: AF1286
6 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF1286
AF1286-SP
Detection of Human Cystatin E/M by Western Blot.
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Product Details
Citations (6)
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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Cystatin E/M Antibody Summary

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 either lyophilized or 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.

Scientific Data

Western Blot Detection of Human Cystatin E/M antibody by Western Blot. View Larger

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.

Simple Western Detection of Human Cystatin E/M antibody by Simple Western<sup>TM</sup>. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.2 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 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

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Citations for Human Cystatin E/M 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.

6 Citations: Showing 1 - 6
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  1. Glycosylation is important for legumain localization and processing to active forms but not for cystatin E/M inhibitory functions
    Authors: NN Lunde, MH Haugen, KB Bodin Lars, I Damgaard, SJ Pettersen, R Kasem, W Rut, M Drag, M Poreba, HT Johansen, R Solberg
    Biochimie, 2017-05-17;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  2. Correction: Nuclear Legumain Activity in Colorectal Cancer
    Authors: Mads H. Haugen, Harald T. Johansen, Solveig J. Pettersen, Rigmor Solberg, Klaudia Brix, Kjersti Flatmark et al.
    PLoS ONE
  3. Nuclear Legumain Activity in Colorectal Cancer
    Authors: Mads H. Haugen, Harald T. Johansen, Solveig J. Pettersen, Rigmor Solberg, Klaudia Brix, Kjersti Flatmark et al.
    PLoS ONE
  4. Differential secretome analysis reveals CST6 as a suppressor of breast cancer bone metastasis.
    Cell Res., 2012-06-12;22(9):1356-73.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells, Whole Tissue
    Applications: IHC, Neutralization, Western Blot
  5. 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, 2010-01-15;10(1):17.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  6. 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., 2008-07-07;88(9):910-25.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC

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