Human SODD/BAG4 Antibody Summary
Accession # O95429
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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 SODD/BAG4 by Western Blot. Western blot shows lysates of MOLT-4 human acute lymphoblastic leukemia cell line, Raji human Burkitt's lymphoma cell line, and A431 human epithelial carcinoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human SODD/BAG4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4350) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). Specific bands were detected for SODD/BAG4 at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
SODD/BAG4 in A431 Human Cell Line. SODD/BAG4 was detected in immersion fixed A431 human epithelial carcinoma cell line using Goat Anti-Human SODD/BAG4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4350) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Western Blot Shows Human SODD/BAG4 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and SODD/BAG4 knockout HeLa cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human SODD/BAG4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4350) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for SODD/BAG4 at approximately 72 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa 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
- 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.
Tumor necrosis factor receptor-1 (TNF-R1) and other TNF receptor superfamily members, such as DR3, contain intracellular death domains (DD) and are capable of initiating apoptosis when activated by their ligands. Silencer of Death Domains (SODD) was identified as being involved in the cellular mechanism to protect against ligand-independent signaling by TNF-R1 and other DD receptors. SODD, also known as Bcl-2-Associated Athanogene 4 (BAG4), is a 457 amino acid (aa), anti‑apoptotic protein that functions through interactions with a variety of proteins including Bcl-2, Raf-protein kinase, steroid hormone receptors, growth factor receptors, and members of the heat shock protein 70 kDa family. SODD is a ubiquitously expressed, cytoplasmic protein that contains a C‑terminal BAG domain that can bind and inhibit the chaperone activity of Hsc70/Hsp70. The association of SODD with the DD of TNF-R1 prevents constitutive activation of the TNF-R1 signaling pathway. Binding of TNF to TNF-R1 releases SODD and permits adapter molecules such as TRADD to associate with TNF-R1 leading to the activation of TNF signaling pathways such as apoptosis and NF kappa B activation.
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