The immunosuppressant cyclosporin A (CsA) forms a trimolecular complex with cyclophilin and calcineurins to inhibit calcineurin phosphatase activity. Cyclophilins are conserved, ubiquitous and abundant cytosolic peptidyl-prolyl cis-trans isomerases that accelerate the isomerization of XaaPro peptide bonds and the refolding of proteins. Human cyclophilin A (CyPA), an intracellular protein of 165 amino acids, is the target of the Cyclosporin A and is encoded by a single unique gene conserved from yeast to humans. CyPA is known for its involvement in T cell differentiation and proliferation and is highly expressed in brain. CyPA is incorporated into the virion of the type 1 human immunodeficiency virus (HIV-1) via a direct interaction with the capsid domain of the viral Gag polyprotein and is crucial for efficient viral replication. Cyclophilin B (CyPB) is a member of the cyclophilin family with specific N- and C-terminal extensions. Unlike CyPA, CyPB has a signal sequence leading to its translocation in the endoplasmic reticulum. CyPB is secreted in biological fluids such as blood or milk and binds to a specific receptor present on the human lymphoblastic cell line Jurkat and on human peripheral blood lymphocytes.
Background References
1. Zhang D et al. Tubular p53 Regulates Multiple Genes to Mediate AKI. J Am Soc Nephrol N/A:N/A (2014).
2. Stocki P et al. Depletion of cyclophilins B and C leads to dysregulation of endoplasmic reticulum redox homeostasis. J Biol Chem 289:23086-96 (2014).
Sequence Similarity
Belongs to the cyclophilin-type PPIase family. PPIase B subfamily.
Western blot analysis of Cyclophilin B on different lysates with Rabbit anti-Cyclophilin B antibody (ET1609-58) at 1/1,000 dilution.
Lane 1: HepG2 cell lysate Lane 2: HeLa cell lysate Lane 3: U-87 MG cell lysate Lane 4: NIH/3T3 cell lysate Lane 5: PC-12 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 24 kDa Observed band size: 20 kDa
Exposure time: 3 minutes; ECL: K1802;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-58) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Knockdown (KD)
Western blot analysis of Cyclophilin B on different lysates with Rabbit anti-Cyclophilin B antibody (ET1609-58) at 1/2,000 dilution.
Lane 1: HAP1-parental cell lysate Lane 2: HAP1-Cyclophilin B KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 24 kDa Observed band size: 20 kDa
Exposure time: 12 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-58) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded rat epididymis tissue using anti-Cyclophilin B antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-58, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human thyroid tissue using anti-Cyclophilin B antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-58, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-Cyclophilin B antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-58, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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