Human Endophilin B1/Bif-1 Antibody

AF7456 Datasheet / COA / SDS

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AF7456
AF7456-SP

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Detection of Human Endophilin B1/Bif‑1 by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Endophilin B1/Bif-1 Antibody Summary

Species Reactivity

Human

Specificity

Detects human Endophilin B1/Bif-1 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human SH3GLB2 is observed.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human Endophilin B1/Bif-1
Ala33-Asn189
Accession # Q9Y371

Formulation

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunohistochemistry

3-15 µg/mL

See below

Immunocytochemistry

5-15 µg/mL

See below

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.

Data Examples

Western Blot

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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.

Immunocytochemistry

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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.

Immunohistochemistry

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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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

Reconstitution Buffer Available

Reconstitution Buffer 1 (PBS)

Shipping

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: 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.

Long Name

SH3-domain GRB2-like Endophilin B1/Bax-interacting Factor 1

Entrez Gene IDs

51100 (Human); 54673 (Mouse); 292156 (Rat)

Alternate Names

Bax-interacting factor 1; Bif1; Bif-1; Bif-1dJ612B15.2; CGI-61; Endophilin B1; endophilin-B1; KIAA0491endophilin B1; SH3 domain-containing GRB2-like protein B1; SH3-containing protein SH3GLB1; SH3-domain GRB2-like endophilin B1; SH3-domain, GRB2-like, endophilin B1; SH3GLB1