Western blot - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

Lanes 1 - 3 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml (5% BSA BLOCK)
Lanes 4 - 6 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml (5% MILK BLOCK)

Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 6 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

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

Performed under reducing conditions.

Predicted band size : 42 kDa
Observed band size : 42 kDa


Exposure time : 8 minutes

Immunofluorescence - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

Immunofluorescence using ab8226 at 5ug/ml incubated for 1 hour on Rat Colon Cancer cells.

Cells were fixed with ice-cold methanol for 5 mins, then for all following steps, permeabilised in TBS-T for 30 mins, blocked with 5% BSA for 30 mins and then washed in TBS-T. Secondary antibody was Alexa Fluor 488 goat anti-mouse IgG at 1/1000 incubated for 1 hour. Cells were counterstained with DAPI. Image at 400X magnification. All incubations were at room temperature.

The beta actin fibres can be seen arrayed around the edge of the cells.

Western blot - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 0.5 µg/ml + HeLa cell lysate

Secondary
Goat polyclonal to mouse IgG H&L (HRP) at 1/5000 dilution

Performed under non-reducing conditions.

Predicted band size : 42 kDa
Observed band size : 42 kDa


Exposure time : 30 seconds

Immunocytochemistry - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

ab8226 staining beta Actin in human AGS cells by Immunocytochemistry. Cells were fixed with formaldehyde, permeabilized with 0.025% Triton-X in TBS and blocking with 5% serum was performed for 1 hour at 23⁰C. Samples were incubated with primary antibody (1/500) for 1 hour at 23°C. An HRP^®-conjugated goat polyclonal to mouse IgG was used undiluted as secondary antibody.

This image is a courtesy of Anonymous Abreview

Immunocytochemistry/ Immunofluorescence - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

ab8226 staining beta Actin in human gastric epithelial AGS cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in formaldehyde. Permeabilization and blocking was carried out using 5% BSA containing 0.025% Triton X in TBS for 1 hour at 23°C. Primary antibody was used at 5µg/ml for 1 hour at 23°C. An Alexa Fluor 488^® conjugated goat polyclonal to mouse IgG was used as the secondary antibody at a 1/1000 dilution.

This image is a courtesy of Anonymous Abreview

Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 42 kDa


Exposure time : 10 seconds

Western blot image using the Optiblot Reducing Electrophoresis Kit - 10 x 10 cm (4-20%) (ab119220) with the
Prism Ultra Protein Ladder (ab116028) 5µl used. We recommend using our ECL substrate kit (ab65623).
Loading Control (ab8226) used at 1 µg/ml.

Lane 1 : Hela cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : A431 cell lysate
Lane 4 : HEK293 cell lysate
Lane 5 : HepG2 cell lysate

Lysates loaded at 20 µg per lane.

Western blot - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)

Lane 1 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution
Lane 2 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/10000 dilution
Lanes 3 - 4 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/500 dilution

Lane 1 : HeLa Cell lysate
Lane 2 : HeLa Cell lysate
Lane 3 : 293 Cell lysate
Lane 4 : 3T3 Mouse Cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
Rabbit polyclonal Secondary Antibody to Mouse IgG - H&L (HRP) (ab6728) at 1/5000 dilution

Performed under reducing conditions.

Predicted band size : 42 kDa


Exposure time : 10 seconds