PIBF1 in Human Breast Cancer Tissue.
PIBF1 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Sheep Anti-Human PIBF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5559) at 3 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). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human PIBF1 by Western Blot.
Western blot shows lysates of JEG‑3 human epithelial choriocarcinoma cell line and human breast cancer tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human PIBF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5559) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for PIBF1 at approximately 98 kDa and 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
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.
PIBF-1 (Progesterone induced blocking factor 1) is a 90-92 kDa progesterone-inducible molecule initially isolated from the lymphocytes of pregnant women. It is produced by lymphocytes plus villous trophoblast cells, and modulates the activity of cytotoxic NK cells. Human PIBF-1 is 757 amino acids (aa) in length. It contains a possible signal sequence (aa 1-26), two Leu-zippers (aa 311-323 and 643-664), two NLSs (aa 282-285 and 567-573), a bZIP sequence (aa 574-614), and an ER retention motif (aa 752-757). PIBF-1 is found in the nucleus, cytoplasm, and circulation, the result of multiple splice variants. SDS-page shows 90 kDa, 80-83 kDa, 48-52 kDa, 34-35 kDa, and 10-12 kDa isoforms. The 90 kDa form is nuclear and full-length, while the 34-35 kDa form is secreted and likely represents aa 1-223 spliced to aa 683-757. There is a 50-55 kDa form that is apparently intracellular, possibly bioactive and likely represents the N-terminus of the molecule. Over aa 19‑419, human PIBF-1 shares 88% aa identity with mouse PIBF-1.
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