Anti-Caveolin-3 antibody - Caveolae Marker

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Caveolin-3 antibody - Caveolae Marker (AB2912)

Immunocytochemistry/Immunofluorescence analysis of 70% confluent log phase A-375 cells. Cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. Samples were incubated with ab2912 at 1μg/ml in 1% BSA for 3 hours at room temperature and then labelled with Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (H+L) at a dilution of 1/2000 for 45 minutes at room temperature (panel a : green). Nuclei (panel b : blue) were stained with DAPI. F-actin (panel c : red) was stained with Alexa Fluor^® 555 Rhodamine Phalloidin (1/300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-Caveolin-3 antibody - Caveolae Marker (AB2912)

Flow cytometry analysis of U-87 MG cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with ab2912 (red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor^® 488-conjugated goat anti-rabbit secondary antibody at a dilution of 1/400 for 30 minutes at room temperature. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Caveolin-3 antibody - Caveolae Marker (AB2912)

Immunocytochemistry analysis of Caveolin-3 in HeLa Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) (right panel) or with ab2912 at a dilution of 1/20 overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Caveolin-3 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Caveolin-3 antibody - Caveolae Marker (AB2912)

Immunocytochemistry analysis of Caveolin-3 in C2C11 Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) (right panel) or with ab2912 at a dilution of 1/20 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Caveolin-3 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

• IP

Supplier Data

Immunoprecipitation - Anti-Caveolin-3 antibody - Caveolae Marker (AB2912)

Caveolin-3 was immunoprecipitated using 5 μg of ab2912 from mouse heart tissue lysate (Lane 3) using the protein A beads. Normal rabbit IgG was used as a isotype control (Lane 2). 10% input represents the cell extract used for immunoprecipitation (Lane 1). Western blot analysis was performed using ab2912 and HRP-conjugated goat anti-rabbit IgG (H+L) at a dilution of 1/2500. Chemiluminescent detection was performed.

All lanes:

Immunoprecipitation - Anti-Caveolin-3 antibody - Caveolae Marker (ab2912)

Predicted band size: 17 kDa

false

• WB

Supplier Data

Western blot - Anti-Caveolin-3 antibody - Caveolae Marker (AB2912)

Blocked with 5% skimmed milk.

All lanes:

Western blot - Anti-Caveolin-3 antibody - Caveolae Marker (ab2912)

Lane 1:

Rat heart tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

HEK293 cell lysate at 20 µg

Lane 4:

Rat skeletal muscle tissue lysate at 20 µg

Lane 5:

Mouse muscle tissue lysate at 20 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG (H+L) at 1/2500 dilution

Predicted band size: 17 kDa

Observed band size: 17 kDa

true

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caveolin-3 antibody - Caveolae Marker (AB2912)

Immunohistochemistry was performed on normal biopsies of deparaffinized mouse lymph node tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/200 with a rabbit polyclonal antibody recognizing Caveolin-3 ab2912 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

• WB

Supplier Data

Western blot - Anti-Caveolin-3 antibody - Caveolae Marker (AB2912)

All lanes:

Western blot - Anti-Caveolin-3 antibody - Caveolae Marker (ab2912)

All lanes:

Rat cardiac muscle tissue lysate

Predicted band size: 17 kDa

false

• WB

CiteAb

Western blot - Anti-Caveolin-3 antibody - Caveolae Marker (AB2912)

Caveolin-3 western blot using anti-Caveolin-3 antibody - Caveolae Marker ab2912. Publication image and figure legend from Niesman, I. R., Schilling, J. M., et al., 2014, J Neuroinflammation, PubMed 24593993.

ab2912 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab2912 please see the product overview.

Caveolin (Cav)-1 knock-out (KO) and Cav-3 KO mice have altered expression and membrane/lipid raft (MLR) localization of key neuronal and glial proteins. Sucrose density fractionation (SDF) and Western blot (WB) of wild-type (WT), Cav-1 KO, and Cav-3 KO brains. Buoyant fractions (BF) contain the cholesterol and sphingolipid enriched MLR, while heavy farctions (HF) contain non-MLR cellular components. (A) WB detection of PSD-95, NR2B, NR1 and TrkB expression in BF and HF. (B) WB detection of toll-like receptor-4 (TLR4), AA2AR, A3AR, and A1AR expression in BF and HF. (C) WB detection of LRP-1 and LDL-R expression in BF and HF. Bottom left, WB analysis of GAPDH in whole cell lysates (WCL) from which SDF were generated for each group. Bottom right, WB shows loss of Cav-1 and Cav-3 protein expression in the transgenic mouse used in the present study.

false