Anti-Clathrin heavy chain antibody [X22]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Clathrin heavy chain antibody [X22] (AB2731)

Immunocytochemistry/Immunofluorescence analysis of Clathrin heavy chain shows staining in U251 cells. Clathrin, Heavy chain staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2731 (1 : 200) over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clathrin heavy chain antibody [X22] (AB2731)

Immunohistochemistry was performed on normal biopsies of deparaffinized Human colon 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 : 100 with a mouse monoclonal antibody recognizing Clathrin Heavy chain ab2731 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Clathrin heavy chain antibody [X22] (AB2731)

Immunocytochemistry/Immunofluorescence analysis of Clathrin heavy chain shows staining in HeLa cells. Clathrin, Heavy chain staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2731 (1 : 200) over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Clathrin heavy chain antibody [X22] (AB2731)

Immunocytochemistry/Immunofluorescence analysis of Clathrin heavy chain shows staining in NCI-H460 cells. Clathrin, Heavy chain staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2731 (1 : 200) over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clathrin heavy chain antibody [X22] (AB2731)

Immunohistochemistry was performed on cancer biopsies of deparaffinized Human breast carcinoma 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 : 100 with a mouse monoclonal antibody recognizing Clathrin Heavy chain ab2731 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-Clathrin heavy chain antibody [X22] (AB2731)

All lanes:

Western blot - Anti-Clathrin heavy chain antibody [X22] (ab2731) at 1/300 dilution

All lanes:

Human brain lysates at 25 µg

Secondary

All lanes:

HRP-conjugated goat anti-mouse IgG + IgM (H+L)

Predicted band size: 191 kDa

true

• WB

Unknown

Western blot - Anti-Clathrin heavy chain antibody [X22] (AB2731)

All lanes:

Western blot - Anti-Clathrin heavy chain antibody [X22] (ab2731) at 1/500 dilution

Lane 1:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 191 kDa

Observed band size: 180 kDa,240 kDa,450 kDa

false

• ICC/IF

AbReview29224****

Immunocytochemistry/ Immunofluorescence - Anti-Clathrin heavy chain antibody [X22] (AB2731)

Xenopus laevis cytoplasmic egg extract visualized live with primary and secondary antibody addition [red is anti-Clathrin heavy chain X22 (ab2731) with goat anti-mouse Alexa Fluor 568 secondary, green is anti-HIP1R (ab77297) with goat anti-rabbit Alexa Fluor 488 secondary]. Large red structures are probably aggregates, but the small structures appear to be specific for vesicle staining.