Anti-Caveolin-1 antibody - Caveolae Marker

• IP

Unknown

Immunoprecipitation - Anti-Caveolin-1 antibody - Caveolae Marker (AB18199)

Caveolin 1 - Caveolae Marker was immunoprecipitated using 0.5mg Mouse Heart whole tissue lysate, 5µg of Rabbit polyclonal to Caveolin 1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Heart whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab18199.
Secondary : Clean blot (HRP conjugate) at 1/1000 dilution.
Band : 20kDa : Caveolin 1 - Caveolae Marker.

All lanes:

Immunoprecipitation - Anti-Caveolin-1 antibody - Caveolae Marker (ab18199)

Predicted band size: 20 kDa

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• WB

Lab

Western blot - Anti-Caveolin-1 antibody - Caveolae Marker (AB18199)

Lanes 1 - 4 : Merged signal (red and green). Green - ab18199 observed at 21-24 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

ab18199 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 3% milk in TBS-T (0.1% Tween^®) before incubation with ab18199 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Caveolin-1 antibody - Caveolae Marker (ab18199) 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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• WB

Unknown

Western blot - Anti-Caveolin-1 antibody - Caveolae Marker (AB18199)

ab18199 recognizes the full length tagged recombinant Caveolin 1 protein (ab114170) which has an expected molecular weight of 46 kDa.

All lanes:

Western blot - Anti-Caveolin-1 antibody - Caveolae Marker (ab18199) at 1 µg/mL

All lanes:

Western blot - Recombinant Human Caveolin-1 protein (<a href='/en-us/products/proteins-peptides/recombinant-human-caveolin-1-protein-ab114170'>ab114170</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 20 kDa

true

Exposure time: 1min

• WB

Unknown

Western blot - Anti-Caveolin-1 antibody - Caveolae Marker (AB18199)

ab18199 recognizes the full length tagged recombinant Caveolin 1 protein (ab114170) which has an expected molecular weight of 46 kDa.

All lanes:

Western blot - Anti-Caveolin-1 antibody - Caveolae Marker (ab18199) at 1 µg/mL

All lanes:

Western blot - Recombinant Human Caveolin-1 protein (<a href='/en-us/products/proteins-peptides/recombinant-human-caveolin-1-protein-ab114170'>ab114170</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 20 kDa

true

Exposure time: 1min

• WB

Project901****

Western blot - Anti-Caveolin-1 antibody - Caveolae Marker (AB18199)

All lanes:

Western blot - Anti-Caveolin-1 antibody - Caveolae Marker (ab18199) at 1 µg/mL

Lane 1:

Mouse Heart Lysate at 10 µg

Lane 2:

Human Heart Lysate at 10 µg

Secondary

All lanes:

Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/5000 dilution

Predicted band size: 20 kDa

Observed band size: 20 kDa

false

• WB

Project901****

Western blot - Anti-Caveolin-1 antibody - Caveolae Marker (AB18199)

All lanes:

Western blot - Anti-Caveolin-1 antibody - Caveolae Marker (ab18199) at 1 µg/mL

Lane 1:

Western blot - NIH/3T3 whole cell lysate (<a href='/en-us/products/cell-lysates/nih-3t3-whole-cell-lysate-ab7179'>ab7179</a>) at 10 µg

Lane 2:

MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Lane 3:

Heart (Mouse) Tissue Lysate at 10 µg

Lane 4:

Heart (Rat) Tissue Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 20 kDa

Observed band size: 20 kDa

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