S100A4 Antibody (3352C) [Unconjugated]

S100A4 was detected in perfusion fixed paraffin-embedded sections of mouse Lung using Rabbit Anti-Mouse S100A4, Monoclonal Antibody (Catalog # MAB11727) at 0.1ug/mL at 37° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the nucleus and cytoplasm. Protocol available in COMET™ Panel Builder.

S100A4 was detected in immersion fixed paraffin-embedded sections of mouse Spleen using Rabbit Anti-Mouse S100A4, Monoclonal Antibody (Catalog # MAB11727) at 0.1ug/mL at 37° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the nucleus and cytoplasm. Protocol available in COMET™ Panel Builder.

S100A4 was detected in perfusion fixed paraffin-embedded sections of mouse spleen using Rabbit Anti-Mouse S100A4 Monoclonal Antibody (Catalog # MAB11727) at 0.05 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003) or the HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the nucleus and cytoplasm. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

S100A4 was detected in perfusion fixed paraffin-embedded sections of mouse thymus using Rabbit Anti-Mouse S100A4 Monoclonal Antibody (Catalog # MAB11727) at 0.05 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003) or the HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the nucleus and cytoplasm. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

S100A4 was detected in Immersion fixed RAW264.7 cells (Positive) and low in Neuro2A cells (Low expression) using Rabbit Anti-Mouse S100A4 Monoclonal Antibody (Catalog # MAB11727) at 3 µg/ml for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to the nucleus and cytoplasm of RAW264.7 cells. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Western Blot shows lysates of NIH‑3T3 mouse embryonic fibroblast cell line. PVDF membrane was probed with 0.05 µg/ml of Rabbit Anti-Mouse S100A4 Monoclonal Antibody (Catalog # MAB11727) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for S100A4 at approximately 11 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Left: Simple Western lane view shows lysates of C2C12 mouse myoblast cell line, loaded at 0.2 mg/ml. A specific band was detected for S100A4 at approximately 9 kDa (as indicated) using both 1 µg/ml and 5 µg/ml of Rabbit Anti-Mouse S100A4 Monoclonal Antibody (Catalog # MAB11727) followed by HRP-conjugated Goat Anti-Rabbit Secondary Antibody (Catalog # 042-206). This experiment was conducted under reducing conditions and using the 2-40kDa separation system. Right: Simple Western electropherogram showing the same Rabbit Anti-Mouse S100A4 Monoclonal Antibody (Catalog # MAB11727) tested at 1 µg/ml (blue line) and 5 µg/ml (green line) in the C2C12 mouse myoblast cell line.

• Reactivity: Mu
• Applications: WB, Simple Western, IHC, ICC/IF, mIF
• Clone: 3352C
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

S100A4 Antibody (3352C) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: E. coli-derived mouse S100A4
  Leu20-Pro219
  Accession # P07091
• Specificity: Detects recombinant mouse S100A4 protein in Direct ELISA.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 0.3-3 ug/mL
  • Immunohistochemistry 0.05-10 ug/mL
  • Multiplex Immunofluorescence 0.1 ug/mL
  • Simple Western 1-5 ug/mL
  • Western Blot 0.05 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Reconstitution Instructions: Reconstitute lyophilized material at 0.2 mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for S100A4 Antibody (3352C) [Unconjugated]

• 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

Background

Mouse S100 calcium-binding protein A4 (S100A4; also metastasin, mts1, metastatic cell protein, and placental calcium-binding protein) is a 12 kDa member of the S100 family of calcium-binding proteins. S100A4 is 101 amino acids (aa) in length and contains two calcium-binding EF-hand domains (aa 12 - 47 and 50 - 85). Mouse S100A4 shares 96%, 93%, and 90% aa sequence identity with rat, human, and bovine S100A4, respectively. S100A4 is involved in regulating the metastatic behavior of tumor cells. It is ubiquitously expressed in tumor cells and normal cells, but the level of S100A4 expression correlates with the metastatic potential of tumor cells.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.