Detects human Semaphorin 4C in direct ELISAs and Western blots. In direct ELISAs, approximately 50% cross-reactivity with recombinant mouse (rm) Sema4C is observed, and approximately 10% cross-reactivity with recombinant human (rh) Sema4G is observed, and less than 5% cross-reactivity with rhSema3B and rmSema3F is observed.
Polyclonal Sheep IgG
Mouse myeloma cell line NS0-derived recombinant human Semaphorin 4C Ala21-Gly663 Accession # Q9C0C4
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 Human Semaphorin 4C by Western Blot.
Western blot shows lysates of SW13 human adrenal cortex adenocarcinoma cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Semaphorin 4C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6125) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Semaphorin 4C at approximately 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Semaphorin 4C in Human Brain.
Semaphorin 4C was detected in immersion fixed paraffin-embedded sections of human brain (cortex) using Sheep Anti-Human Semaphorin 4C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6125) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). 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 neuronal cell bodies. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human Semaphorin 4C by Simple WesternTM.
Simple Western lane view shows lysates of SW13 human adrenal cortex adenocarcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Semaphorin 4C at approximately 121 kDa (as indicated) using 25 µg/mL of Sheep Anti-Human Semaphorin 4C Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6125) 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. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
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.
Background: Semaphorin 4C
Sema4C (Semaphorin 4C; also Sema I and M-SemaF) is a 100-105 kDa, class IV member of the semaphorin family of proteins. It is expressed by precursors to neurons and myocytes, and may regulate their differentiation into mature forms. Mature human Sema4C is a type I transmembrane glycoprotein that is 813 amino acids (aa) in length. It contains a 643 aa extracellular region (aa 21-663) that is characterized by the presence of one Sema domain (aa 30-497), a PSI region (aa 499‑551), and an Ig-like C2-type domain (aa 556-644). The cytoplasmic region interacts with PZD-domain containing proteins. There are three potential splice variants. One demonstrates an alternative start site at Met324, a second shows a deletion of aa 173-211, and a third contains an 89 aa substitution for aa 1-36. Over aa 21‑663, human Sema4C shares 85% aa identity with mouse Sema4C.
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We have 1 review tested in 1 application: Western Blot.