Alexa Fluor® 488 Anti-Caveolin-1 抗体 [EPR15554] - N-terminal

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Caveolin-1 antibody [EPR15554] - N-terminal - Caveolae Marker (AB225380)

Flow cytometry overlay histogram showing wild-type (green line) and CAV1 knockout (ab255371) HeLa cells stained with ab225380 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab225380) (1x 10⁶ in 100μl) at 0.04μg/ml (1/12500) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor 488 ^® (ab199091) used at the same concentration and conditions as the primary antibody (wild-type - black line knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Caveolin-1 antibody [EPR15554] - N-terminal - Caveolae Marker (AB225380)

ab225380 staining Caveolin-1 in A673 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab225380 at 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in A673 cells fixed with 4% formaldehyde (10 min).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Caveolin-1 antibody [EPR15554] - N-terminal - Caveolae Marker (AB225380)

ab225380 staining Caveolin-1 in wild-type HeLa cells (top panel) and CAV1 knockout HeLa cells (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab225380 at 1/500 dilution and ab195884 (Rat monoclonal to Tubulin - Alexa Fluor^® 647) at 1/100 dilution overnight at 4°C. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Caveolin-1 antibody [EPR15554] - N-terminal - Caveolae Marker (AB225380)

Overlay histogram showing A673 cells stained with ab225380 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab225380, 1/5000 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor^® 488 (ab199091) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

This antibody gave a positive signal in A673 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.