Detects human and mouse S100A4 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant mouse (rm) S100A8, rmS100A6, rmS100A9, rmS100A10, rmS100A13, rmS100A16, recombinant human (rh) S100A1, rhS100A7, rhS100A11, rhS100B, and rhS100P is observed.
Polyclonal Sheep IgG
E. coli-derived recombinant mouse S100A4 Ala2-Lys101 Accession # P07091
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.
Detection of Mouse S100A4 by Western Blot.
Western blot shows lysates of 3T3‑L1 mouse embryonic fibroblast adipose‑like cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse S100A4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4138) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for S100A4 at approximately 11 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Detection of Mouse S100A4 by Simple WesternTM.
Simple Western lane view shows lysates of 3T3‑L1 mouse embryonic fibroblast adipose-like cell line, loaded at 0.2 mg/mL. A specific band was detected for S100A4 at approximately 12 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human/Mouse S100A4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4138) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Western Blot Shows Sheep Anti-Human/Mouse S100A4 Antigen Affinity-purified Polyclonal Antibody Specificity by Using Knockout Cell Line.
Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and S100A4 knockout HeLa cell line (KO). PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse S100A4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4138) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for S100A4 at approximately 12 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockoutHeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
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.
S100A4 (also named metastasin, Mtsl and calvasculin) is an 11 kDa member of the S100 (soluble in 100% saturated ammonium sulfate) family of proteins (1‑5). The S100 family is a further classified as a member of the EF-hand superfamily of Ca++-binding proteins. These participate in both calcium‑dependent and calcium‑independent protein-protein interactions. The hallmark of this superfamily is the EF-hand motif that consists of a Ca++-binding site flanked by two alpha -helices (helix E and helix F) that were originally identified in a right-handed model of carp muscle calcium‑binding protein (6). Mouse S100A4 is 101 amino acids (aa) in length (1, 2). It contains two EF hand domains, one between aa 12‑47, and a second between aa 50‑85. The first domain has a 14 aa cation-binding motif and binds Ca++ with low affinity. The second Ca++-binding motif is 12 aa in length and binds Ca++ with high affinity. S100A4 has no classical signal sequence but is secreted from cells (3, 7). Mouse S100A4 shares 93%, 96% and 89% aa identity with human, rat and canine S100A4, respectively. S100A4 exists as dimer (8, 9, 10). Extracellular S100A4 is reported to induce MMP production, activate MMPs, promote neurite outgrowth and stimulate cardiomyocyte proliferation (4, 10, 11, 12, 13). Within the cell, dimers are likely the functional unit. Here, they are constitutive homo- or heterodimers (with S100A1) that interact with Ca++, undergo a conformational change, and subsequently bind to cytoplasmic targets. Known targets include p53, myosin heavy chain II, F-actin and liprin beta 1 (4, 14). In general, it can be said that S100A4 blocks target phosphorylation and multimerization (4, 7, 14). S100A4 activity has been associated with cell transformation. It seems likely this is either coincidental, or a consequence, rather than a cause of transformation (3).
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S100 Calcium Binding Protein A4
Entrez Gene IDs:
6275 (Human); 20198 (Mouse); 24615 (Rat)
18A2; 42A; Calvasculin; CAPL; CAPLS100 calcium binding protein A4 (calcium protein, calvasculin, metastasin; fibroblast-specific protein-1,42A; FSP1; leukemia multidrug resistance associated protein; malignant transformation suppression 1; Metastasin; MTS1; murine placental homolog); P9KA; PEL98; Placental calcium-binding protein; Protein Mts1; protein S100-A4; S100 calcium binding protein A4; S100 calcium-binding protein A4 (calcium protein, calvasculin, metastasin, murine placental homolog); S100 calcium-binding protein A4; S100A4
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