Detects Betacellulin/BTC in direct ELISAs and Western blots.
Polyclonal Goat IgG
E. coli-derived recombinant human Betacellulin Asp32-Tyr111 Accession # P35070
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.
<0.10 EU per 1 μg of the antibody by the LAL method.
Measured by its ability to neutralize Betacellulin/BTC-induced proliferation in the Balb/3T3 mouse embryonic fibroblast cell line. The Neutralization Dose (ND50) is typically 0.04-0.08 µg/mL in the presence of 1 ng/mL Recombinant Human Betacellulin/BTC.
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.
Detection of Human Betacellulin/BTC by Western Blot. Western blot shows lysates of MCF‑7 human breast cancer cell line and OVCAR‑3 human ovarian carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human Betacellulin/BTC Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-261-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Betacellulin/BTC at approximately 32 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Betacellulin/BTC in Human Pancreas. Betacellulin/BTC was detected in immersion fixed paraffin-embedded sections of human pancreas using Goat Anti-Human Betacellulin/BTC Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-261-NA) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to islets. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Betacellulin/BTC in Human Lung Cancer Tissue. Betacellulin/BTC was detected in immersion fixed paraffin-embedded sections of human lung cancer tissue using Goat Anti-Human Betacellulin/BTC Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-261-NA) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Cell Proliferation Induced by Betacellulin/BTC and Neutralization by Human Betacellulin/BTC Antibody. Recombinant Human Betacellulin/BTC (Catalog # 261‑CE) stimulates proliferation in the Balb/3T3 mouse embryonic fibroblast cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human Betacellulin/BTC (1 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human Betacellulin/ BTC Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-261-NA). The ND50 is typically 0.04‑0.08 µg/mL.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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.
Betacellulin (BTC) is a member of the EGF family of cytokines that is comprised of at least ten proteins including EGF, TGF-alpha, amphiregulin, HB-EGF, and the various heregulins. All of these cytokines are synthesized as transmembrane precursors and are characterized by the presence of one or more EGF structural units in their extracellular domain. The soluble forms of these cytokines are released by proteolytic cleavage. BTC, a heparin-binding protein, was originally isolated from the conditioned media of mouse pancreatic beta tumor cells as a 32 kDa glycoprotein composed of 80 amino acid residues. The cDNA encoding human BTC was cloned from a human breast adenocarcinoma cell line (MCF-7) cDNA library. Human and mouse cDNAs encode BTC precursor proteins of 178 and 177 amino acid residues, respectively. At the amino acid sequence level, human BTC precursor protein exhibits 79% identity with that of the mouse BTC precursor. In a mouse cell line transfected with human BTC cDNA, three forms of soluble human BTC have been detected: the glycosylated, intact BTC composed of 80 amino acid residues, a truncated molecule lacking 12 amino acid residues from the amino terminus, and a second truncated molecule lacking 30 amino acid residues from the amino terminus. The biological activities of these BTC forms were shown to be identical. BTC can bind to the EGF receptor and is a potent mitogen for Balb/c 3T3 fibroblasts, retinal pigment epithelial cells and vascular smooth muscle cells.
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