Rabbit immunoglobulin fraction code x0903

Cells were fixed with ice-cold methanol and incubated with 0.5% Triton. Unspecific epitops were blocked using 20% AB serum. Next, cells were incubated with their respective primary antibody or isotype control (mouse monoclonal anti-CXCR4, 1 : 250 dilution; Abcam; rabbit polyclonal anti-CXCR7, 1 : 500 dilution; GeneTex; mouse IgG1k (MOPC-21); 2 μg ml⁻¹; Abcam; rabbit immunoglobulin fraction (Code X0903) 2 μg ml⁻¹; Dako). After 45 min of incubation, cells were washed and treated with either anti-mouse- or anti-rabbit-Alexa Fluor 488 (10 μg ml⁻¹; Thermo Fisher). Nuclear staining was achieved using DAPI (200 ng ml⁻¹). After fixation with 1% PFA, the cells were visualised using a fluorescence microscope at × 400 magnification (Zeiss Axioplan 2, Carl Zeiss).

Cells were cultivated in 6-well chamber slides and fixated with ice-cold methanol. After treatment with 0.5 % Triton the cells were blocked with 20 % AB serum and incubated with their respective primary antibody and isotype control (mouse monoclonal anti-CXCR4, 1:250 dilution; Abcam, Cambridge, UK; rabbit polyclonal anti-CXCR7, 1:500 dilution; GeneTex, Irvine, CA, USA; mouse IgG1k (MOPC-21); 2 µg/ml; Abcam, Cambridge, UK; rabbit immunoglobulin fraction (Code X0903) 2 µg/ml; Dako, Glostrup, Denmark). After 45 minutes of incubation, cells were washed and incubated with anti-mouse- or anti-rabbit-Alexa Fluor® 488 (10 µg/ml; Thermo Fisher, MA, USA), respectively. Nuclear staining was carried out with DAPI (200 ng/ml). After fixating the cells with 1 % PFA, visualization was achieved using a fluorescence microscope at 400x magnification (Zeiss Axioplan 2, Carl Zeiss, Göttingen, Germany).