PBS, 25% Glycerol, 1% BSA, 0.3% Proclin 300
Isotype control antibodies, to estimate the nonspecific binding of target. Use at concentrations comparable to those of the specific antibody of interest.
Flow cytometric analysis of Ramos (Human Burkitt's lymphoma B lymphocyte) labelling Alexa Fluor® 555 Rabbit IgG Isotype Control Antibody at 1/20 dilution (1μg) / (Black) compared with CD38 antibody / (Red), and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 555 was used as the secondary antibody.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) labelling Alexa Fluor® 555 Rabbit IgG Isotype Control Antibody at 1/20 dilution (1μg) / (Black) compared with β-tubulin (Alexa Fluor® 555 Conjugate) antibody at 1/200 (1 μg) dilution (S0B0135) / (Red) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
ICC shows negative staining in HeLa cells. Alexa Fluor® 555 Rabbit IgG Isotype Control Antibody was used at 1/5 dilution and incubated overnight at 4°C. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue).
ICC shows negative staining in NIH/3T3 cells. Alexa Fluor® 555 Rabbit IgG Isotype Control Antibody was used at 1/5 dilution and incubated overnight at 4°C. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue).
IF shows negative staining in paraffin-embedded human prostatic cancer. Alexa Fluor® 555 Rabbit IgG Isotype Control Antibody was used at 1/5 dilution and incubated overnight at 4°C. Counterstained with DAPI (Blue). Heat mediated antigen retrieval with EDTA buffer pH9.0 was performed before commencing with IF staining protocol.
IF shows negative staining in paraffin-embedded human breast cancer. Alexa Fluor® 555 Rabbit IgG Isotype Control Antibody was used at 1/5 dilution and incubated overnight at 4°C. Counterstained with DAPI (Blue). Heat mediated antigen retrieval with EDTA buffer pH9.0 was performed before commencing with IF staining protocol.
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