Human S100A9 Antibody

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  • Species Reactivity
    Human
  • Specificity
    Detects human S100A9 in direct ELISAs and Western blots. In direct ELISAs, less than 5% cross-reactivity with recombinant mouse S100A9 is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human S100A9
    The2-Pro114
    Accession # P06702
  • 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
Product Datasheets

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Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.2 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunocytochemistry
    1-25 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
  • Knockout Validated
    S100A9 is specifically detected in MDA‑MB‑468 human breast cancer parental cell line but is not detectable in S100A9 knockout MDA‑MB‑468 cell line.
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 S100A9 by Western Blot. Western blot shows lysates of human peripheral blood mononuclear cells (PBMC), human spleen tissue, human tonsil tissue, and human cartilage tissue. PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5578) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for S100A9 at approximately 14 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
S100A9 in MDA‑MB‑468 Human Cell Line. S100A9 was detected in immersion fixed MDA‑MB‑468 human breast cancer cell line using Sheep Anti-Human S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5578) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunohistochemistry
S100A9 in Human Cartilage. S100A9 was detected in immersion fixed paraffin-embedded sections of human cartilage using Sheep Anti-Human S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5578) at 1 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm of chondrocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Intracellular Staining by Flow Cytometry
Detection of S100A9 in Human PBMCs by Flow Cytometry. Human peripheral blood monocytes (PBMC) were stained with Mouse Anti-Human CD14 PE-conjugated Monoclonal Antibody (Catalog # FAB3832P) and either (A) Sheep Anti-Human S100A9 Polyclonal Antibody (Catalog # AF5578) or (B) Sheep IgG Isotype Control (Catalog # 5-001-A) followed by anti-Sheep IgG APC-conjugated Secondary Antibody (Catalog # F0127). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer (Catalog # FC005). View our protocol for Staining Membrane-associated Proteins.
Knockout Validated
Western Blot Shows Human S100A9 Specificity by Using Knockout Cell Line. Western blot shows lysates of MDA‑MB‑468 human breast cancer parental cell line and S100A9 knock out MDA‑MB‑468 cell line (KO). PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5578) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for S100A9 at approximately 14 kDa (as indicated) in the parental MDA‑MB‑468 cell line, but is not detectable in knockout MDA‑MB‑468 cell line. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Knockout Validated
S100A9 Specificity is Shown by Immunocytochemistry in Knockout Cell Line. S100A9 was detected in immersion fixed MDA‑MB‑468 human breast cancer cell line but is not detected in S100A9 knockout (KO) MDA-MB-468 cell line using Sheep Anti-Human S100A9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5578) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
    Reconstitution Buffer Available
  • 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: S100A9

Human S100A9 (also MRP-14, Calgranulin-B, and p14) is a 14 kDa member of the S100 family of EF-hand calcium-binding proteins.  It is 114 amino acids (aa) in length and contains short sequential modules.  There is an N-terminal helical region, followed by a calcium-binding EF-hand domain, two more helical regions, a second EF-hand domain, and three additional helical regions.  S100A9 will noncovalently heterodimerize with S100A8.  In the presence of calcium, this heterodimer will form a heterotetramer. S100A9 is expressed in granulocytes, monocytes, and macrophages during acute and chronic inflammation.  Human S100A9 shares 62% and 57% aa identity with rat and mouse S100A9, respectively.

  • Long Name:
    S100 Calcium Binding Protein A9
  • Entrez Gene IDs:
    6280 (Human); 20202 (Mouse)
  • Alternate Names:
    CAGBMigration inhibitory factor-related protein 14; Calgranulin B; calgranulin-B; Calprotectin L1H subunit; CFAGMRP-14,60B8AG; CGLB; L1AG; LIAG; MAC387; MIF; MRP-14; MRP14Leukocyte L1 complex heavy chain; NIF; P14; protein S100-A9; S100 calcium binding protein A9 (calgranulin B); S100 calcium binding protein A9; S100 calcium-binding protein A9 (calgranulin B); S100 calcium-binding protein A9; S100A9
Related Research Areas

FAQs

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Average Rating: 4 (Based on 1 review)

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We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: ELISA.

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Images Ratings Applications Species Reviewed By Date Details
ELISA Human S100A9 Antibody AF5578 Array AF5578
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Very Good
 ELISA Human Anonymous 01/05/2018
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ELISA Human S100A9 Antibody AF5578
ELISA: Human S100A9 Antibody [AF5578].

Summary

Application ELISA
Sample Tested Serum and Plasma
Species Human

Other Experimental Details

Other Experimental Details This antibody was used as both capture and detection in an ELISA reaction measuring S100A9 in human samples.

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