Human Cystatin E/M Antibody

(1 citations)   
  • Species Reactivity
    Human
  • Specificity
    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.
  • Source
    Monoclonal Mouse IgG2B Clone # 211515
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • 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
    2 µg/mL
    See below
  • Immunohistochemistry
    8-25 µg/mL
    See below
  • 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
  • Neutralization
    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. 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 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.
Immunohistochemistry
Cystatin E/M in Human Skin.

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
  • Reconstitution
    Reconstitute at 0.5 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.

1 Citations: Showing 1 - 1

  1. Activator protein-1 activity regulates epithelial tumor cell identity.
    Authors: Gerdes MJ, Myakishev M, Frost NA, Rishi V, Moitra J, Acharya A, Levy MR, Park SW, Glick A, Yuspa SH, Vinson C
    Cancer Res., 2006;66(15):7578-88.
    Species: Human
    Sample Type: Tissue Homogenates
    Application: WB
Cell and Tissue Staining Kits
Description Application Cat# Citations Images  

Anti-Mouse HRP-DAB Cell & Tissue Staining Kit

CTS002 14
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ExactaChIP Chromatin Immunoprecipitation Kits
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ExactaChIP Chromatin IP Buffer Panel

ChIP ECP001 2
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Immunohistochemistry Reagents
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Antigen Retrieval Reagent-Universal

CTS015 10
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Antigen Retrieval Reagent-Basic

CTS013 4
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Mounting Medium

CTS011 4
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Antigen Retrieval Reagent Sampler (50 mL each)

CTS016
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Antigen Retrieval Reagent-Acidic

CTS014
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NorthernLights Guard Mounting Media

NL996
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VisUCyte Antigen Retrieval Reagent-Universal

VCTS023
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VisUCyte Antigen Retrieval Reagent-Basic

VCTS021
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VisUCyte Antigen Retrieval Reagent-Acidic

VCTS022
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Isotype Controls
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Mouse IgG2B Isotype Control

Ctrl MAB004 32  
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Mouse IgG2B Isotype Control

Ctrl MAB0041 7  
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Mouse IgG2B Isotype Control

Ctrl MAB0042 6
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Secondary Antibodies
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Mouse IgG HRP-conjugated Antibody

WB HAF007 26  
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Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0102B 10  
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Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Flow, IHC, ICC NL007 9
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Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
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Mouse IgG HRP-conjugated Antibody

WB HAF018 9  
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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL008 8
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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 2  
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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL009 5
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse IgG Antibody

WB AF007 4
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Rat Anti-Mouse IgG2B APC-conjugated Antibody

Flow F0133 1  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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Rat Anti-Mouse IgG2B PE-conjugated Antibody

Flow F0132  
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Rat Anti-Mouse IgG2B Fluorescein-conjugated Antibody

Flow F0134 1
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Mouse IgG2B Antibody

Flow MAB0043  
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