ab238462)

ab238462 (red line). The cells were incubated in 1x PBS containing 10 % rat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab238462) (1x^{ab150117) was used at 1/2000 for 30 min on ice.}

ab170190) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab238462).

IHC image of CD147 staining in a section of frozen normal rat colon*.

The section was fixed using 10% formaldehyde in 1XPBS for 10 minutes. No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab238462 (shown in green) at 5g/ml.

The section was then incubated with ab150117 (Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 488, 1/1000)) presabsorbed for 1 hour at room temperature. The secondary-only control image is taken from an identical assay without primary antibody. The section was then mounted using Fluoromount^®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).For IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antibody concentrations and incubation times.

*Tissue obtained from Charles River.