Plays an essential role in cytoplasmic secondary envelopment during viral egress. Interacts with the capsid via the large tegument protein/LTP and participates in its transport to the host trans-Golgi network (TGN) where secondary envelopment occurs. Modulates tegumentation and capsid accumulation at the viral assembly complex.
Western blot analysis of BOLF1 on different lysates with Rabbit anti-BOLF1 antibody (HA723040) at 1/2,000 dilution.
Lane 1: HK1 cell lysate Lane 2: HK1 transfected with BOLF1 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 133 kDa Observed band size: 133 kDa
Exposure time: 4 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 (HA723040) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Cell treatment (CT)
Immunocytochemistry analysis of HK1 cells transfected with BOLF1 labeling BOLF1 with Rabbit anti-BOLF1 antibody (HA723040) at 1/1,000 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-BOLF1 antibody (HA723040) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
☑ Cell treatment (CT)
Immunohistochemical analysis of paraffin-embedded HK1 transfected with BOLF1 cells with Rabbit anti-BOLF1 antibody (HA723040) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723040) at 1/5,000 dilution for 1 hour 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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