Rabbit anti human pu 1 h 135

Decidual cells on cytospin slides were fixed with 4% PFA and permeabilized with 0.1% saponin in PBS for 10 min. The slides were incubated with mouse anti-human CK-7 (1:100; Abcam) and rabbit anti-human PU.1 H-135 (1:600, Santa Cruz). After washing with PBS, the slides were incubated with goat-anti-mouse Cy3 (1:200) and goat-anti-rabbit FITC (3:100) conjugates.

Decidual cells on cytospin slides were fixed with 4% PFA and permeabilized with 0.1% saponin in PBS for 10 min. The slides were incubated with mouse anti-human C1q (1:200, Abcam) and rabbit anti-human PU.1 H-135 (1:600, Santa Cruz). After washing with PBS, the slides were incubated with goat-anti-mouse Cy3 (1:200) and goat-anti-rabbit FITC (3:100) conjugates. As controls, we analyzed the expression of the same in RAW cells using identical conditions.
All primary and secondary antibodies were diluted in 5% BSA and incubated 1 h in ice. The slides were finally mounted using Vectashield DAPI (Vectastain, Vector Laboratories). The cellular localizations of various proteins were examined using Zeiss Axiolmager Z1 fluorescent photomicroscope. The images were captured and recorded using digital camera (Zeiss Axiocam MRc). Cells incubated with Isotype IgG were used as non-specific controls.

Decidual stromal cells on cytospin slides were incubated with mouse anti-human vimentin (1:150; Dako). After washing in PBS, slides were fixed in 4% PFA, and permeabilized in 0.1% saponin in PBS for 10 min. The slides were incubated with rabbit anti-human PU.1 H-135 (1:600; Santa Cruz). After washing with PBS, the slides were incubated with goat-anti-mouse FITC (3:100) and goat-anti-rabbit Cy3 (1:200) conjugates.