Human Endophilin B1/Bif-1 Antibody Summary
Accession # Q9Y371
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.
Endophilin B1/Bif‑1 in Human Pancreatic Cancer Tissue. Endophilin B1/Bif-1 was detected in immersion fixed paraffin-embedded sections of human pancreatic cancer tissue using Sheep Anti-Human Endophilin B1/Bif-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7456) at 3 µg/mL overnight at 4 °C. 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 cytoplasm in epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human Endophilin B1/Bif‑1 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line, Saos-2 human osteosarcoma cell line, and U-118-MG human glioblastoma/astrocytoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Endophilin B1/Bif-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7456) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Endophilin B1/Bif-1 at approximately 43 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Endophilin B1/Bif‑1 in Jurkat Human Cell Line. Endophilin B1/Bif-1 was detected in immersion fixed Jurkat human acute T cell leukemia cell line using Sheep Anti-Human Endophilin B1/Bif-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7456) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
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.
Background: Endophilin B1/Bif-1
Endophilin-B1, also known as SH3GLB1 (Src-Homology 3 Domain-containing GRB2-like protein B1) and BIF-1, is a 40 kDa protein that belongs to the endophilin family of molecules. It is a cytoplasmic and Golgi membrane protein that is expressed in neurons, striated (skeletal and cardiac) muscle cells, and placenta. Endophilin-B1 plays a role in cell homeostasis. In neurons, Endophilin-B1 is phosphorylated by Cdk5, inducing homodimerization and autophagosome formation. In addition, it appears to play a role in the maintenance of mitochondrial integrity. Endophilin-B1 appears to cycle on-and-off the outer mitochondrial membrane (OMM), contributing to OMM integrity. And after the initiation of endocytosis, it also directs EEA1+ TrkA-containing endosomes back into the cell membrane for reuse. Human Endophilin‑B1 is 365 amino acids (aa) in length. It possesses an N-terminal lipid-binding BAR (Bin/Amphiphysin/Rvs) domain (aa 27-261) that contains an internal coiled-coil region, and a C-terminal SH3 domain that binds Pro-rich sequences on potential ligands (309-363). There are at least three potential splice variants for Endophilin-B1. Two are 42 and 43 kDa in size, and possess a 21 and 29 aa insertion after Ser190, respectively. A third potentially utilizes an alternative start site at Met101. Over aa 33-189, human Endophilin-B1 shares 98% aa sequence identity with mouse Endophilin-B1.
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