|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.|
|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.|
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.
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Western Blot: Human S100A8 Antibody [AF4570]
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