Human S100A8 Antibody

(1 Review)
  
  • Species Reactivity
    Human
  • Specificity
    Detects human S100A8 in direct ELISAs and Western blots. In these formats, less than 1% cross-reactivity with recombinant mouse S100A8, recombinant human (rh) S100A1, and rhS100P is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human S100A8
    Met1-Glu93
    Accession # P05109
  • 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
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    See below
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 S100A8 by Western Blot. Western blot shows lysates of human brain tissue, K562 human chronic myelogenous leukemia cell line, and HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human S100A8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4570) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for S100A8 at approximately 10 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Immunocytochemistry
S100A8 in HeLa Human Cell Line. S100A8 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human S100A8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4570) at 10 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.
Intracellular Staining by Flow Cytometry
Detection of S100A8 in HeLa Human Cell Line by Flow Cytometry. HeLa human cervical epithelial carcinoma cell line was stained with Sheep Anti-Human S100A8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4570, filled histogram) or control antibody (Catalog # 5-001-A, open histogram), followed by NorthernLights™ 637-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # NL011). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
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: S100A8

S100A8 (also known as MRP8 and calgranulin A) is a 10 kDa member of the S100 family, EF-hand superfamily of Ca-binding proteins. It is produced by neutrophils and monocytes, and forms Ca‑dependent heterodimer/heterotetramer complexes (termed calprotectin) with S100A9. It functions both intracellularly and extracellularly, where it binds to RAGE and CD36. Human S100A8 is 93 amino acids (aa) in length. It contains two EF-hand motifs (aa 12-47 and 46-81) and one high-affinity Ca‑binding site (aa 59-70). There may be one splice form that shows a 15 aa substitution for the C-terminal 14 amino acids. Although mouse S100A8 is cleaved by MMP‑2 after Asn21, it is unclear if human S100A8 is susceptible. Full-length human S100A8 is 57% and 74% aa identical to mouse and canine S100A8, respectively.

  • Long Name:
    S100 Calcium Binding Protein A8
  • Entrez Gene IDs:
    6279 (Human); 20201 (Mouse)
  • Alternate Names:
    60B8AG; CAGACP-10; Calgranulin A; calgranulin-A; Calprotectin L1L subunit; CFAG; CFAGL1Ag; CGLA; Cystic fibrosis antigen; Leukocyte L1 complex light chain; MA387; MIF; Migration inhibitory factor-related protein 8; MRP8; MRP-8; MRP8S100 calcium binding protein A8 (calgranulin A); NIF; P8; protein S100-A8; S100 calcium binding protein A8; S100 calcium-binding protein A8 (calgranulin A); S100 calcium-binding protein A8; S100A8; Urinary stone protein band A
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Average Rating: 5 (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: Western Blot.

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Western Blot Human S100A8 Antibody AF4570
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 WB Human Anonymous 07/01/2016
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Western Blot Human S100A8 Antibody AF4570
Western Blot: Human S100A8 Antibody [AF4570]

Summary

ApplicationWestern Blot
Sample TestedRecombinant protein
SpeciesHuman

Other Experimental Details

Other Experimental DetailsS100A8 Antibody tested against different concentrations of human recombinant S100A8/A9.

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