Anti-Caveolin-1 antibody [7C8] - Caveolae Marker

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Caveolin-1 antibody [7C8] - Caveolae Marker (AB17052)

ab17052 staining Caveolin-1 in wild-type HeLa cells (top panel) and CAV1 knockout HeLa cells (ab255371) (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 ab17052 at 1/500 dilution and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor^® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor^® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

• ICC

AbReview18155****

Immunocytochemistry - Anti-Caveolin-1 antibody [7C8] - Caveolae Marker (AB17052)

ab17052 staining Caveolin-1 in human Hacat keratinocyte cells by Immunocytochemistry/ Immunofluorescence. The cells were formaldehyde fixed, permeabilised in 0.1% Triton X-100 and then blocked using 1% serum for 1 hour at 25°C. Samples were then incubated with primary antibody at 1/75 for 24 hours at 4°C. The secondary antibody used was conjugated to Alexa Fluor^® 488 (green) used at a 1/500 dilution.

This image is courtesy of an anonymous abreview.

• ICC

AbReview43982****

Immunocytochemistry - Anti-Caveolin-1 antibody [7C8] - Caveolae Marker (AB17052)

Immunocytochemical analysis of Mouse Bend.3 cells, labeling Caveolin-1 with ab17052. Cells were paraformaldehyde fixed, permeabilized with PBS + 0.1% Triton (PBT), and blocked with 10% serum for 1 hour at 22°C. Staining with ab17052 (diluted 1/200) was for 16 hours at 4°C.

This image is courtesy of an anonymous Abreview.

• WB

Lab

Western blot - Anti-Caveolin-1 antibody [7C8] - Caveolae Marker (AB17052)

Lanes 1 - 4 : Merged signal (red and green). Green - ab17052 observed at 21-24 kDa. Red - loading control, ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

ab17052 was shown to react with Caveolin-1 in wild-type A431 cells in western blot. Loss of signal was observed when CAV1 knockout sample was used. Wild-type and CAV1 knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in non-mammalian (TBS-based) blocking solution before incubation with ab17052 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Caveolin-1 antibody [7C8] - Caveolae Marker (ab17052) at 1 µg/mL

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

CAV1 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CAV1 (Caveolin-1) knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-cav1-caveolin-1-knockout-a-431-cell-line-ab269583'>ab269583</a>)

Lane 3:

A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 20 kDa

Observed band size: 21-24 kDa

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• ICC

AbReview4874****

Immunocytochemistry - Anti-Caveolin-1 antibody [7C8] - Caveolae Marker (AB17052)

ab17052 at a 1/500 dilution staining hamster CHO cells by ICC/IF. The cells were paraformaldehyde fixed and blocked with 10% serum prior to incubation with the antibody. Bound antibody was detected using a FITC conjugated goat anti-mouse antibody.

This image is courtesy of an anonymous Abreview