Anti-Actin antibody [IGX3831H] - Loading Control

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Actin antibody [IGX3831H] - Loading Control (AB213251)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized, PBS washing, and blocked with 10% normal goat serum in PBS HeLa (human cervical cancer cells) cells labelling Actin with ab213251 at 1 : 400 (1.20 µg/ml) dilution, followed by ab325309 Alpaca multiclonal [2F3 / 2E5] Anti-human IgG1/IgG4 antibody (ATTO 488) at 1/500 (0.15 µg/ml per sdAb) dilution (green).

Confocal image showing cytoplasmic staining in HeLa cells (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a fluorescence microscope (Zeiss Axio Imager Z1).

Secondary antibody only control : Secondary antibody is ab325309 Alpaca multiclonal [2F3 / 2E5] Anti-human IgG1/IgG4 antibody (ATTO 488) at 1/500 (0.15 µg/ml per sdAb) dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Actin antibody [IGX3831H] - Loading Control (AB213251)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized, PBS washing, and blocked with 10% normal goat serum in PBS HeLa (human cervical cancer cells) cells labelling Actin with ab213251 at 1 : 400 (1.20 µg/ml) dilution, followed by ab325308 Alpaca multiclonal [2F3 / 2E5] Anti-human IgG1/IgG4 antibody (Alexa Fluor® 647) at 1/500 (0.15 µg/ml per sdAb) dilution (magenta).

Confocal image showing cytoplasmic staining in HeLa cells (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a fluorescence microscope (Zeiss Axio Imager Z1).

Secondary antibody only control : Secondary antibody is ab325308 Alpaca multiclonal [2F3 / 2E5] Anti-human IgG1/IgG4 antibody (Alexa Fluor® 647) at 1/500 (0.15 µg/ml per sdAb) dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Actin antibody [IGX3831H] - Loading Control (AB213251)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized, PBS washing, and blocked with 10% normal goat serum in PBS HeLa (human cervical cancer cells) cells labelling Actin with ab213251 at 1 : 400 (1.20 µg/ml) dilution, followed by ab325307 Alpaca multiclonal [2F3 / 2E5] Anti-human IgG1/IgG4 antibody (AZdye 568) at 1/500 (0.15 µg/ml per sdAb) dilution (red).

Confocal image showing cytoplasmic staining in HeLa cells (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a fluorescence microscope (Zeiss Axio Imager Z1).

Secondary antibody only control : Secondary antibody is ab325307 Alpaca multiclonal [2F3 / 2E5] Anti-human IgG1/IgG4 antibody (AZdye 568) at 1/500 (0.15 µg/ml per sdAb) dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Actin antibody [IGX3831H] - Loading Control (AB213251)

ab213251 staining Actin in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), 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 overnight at +4°C with ab213251 at a 5μg/ml concentration, then detected with a goat anti-human (Alexa Fluor® 488) secondary antibody at a 1/2000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Actin antibody [IGX3831H] - Loading Control (AB213251)

ab213251 staining Actin in NIH3T3 cells. The cells were fixed with 4% formaldehyde (10 min), 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 overnight at +4°C with ab213251 at a 5μg/ml concentration, then detected with a goat anti-human (Alexa Fluor^® 488) secondary antibody at a 1/2000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• WB

Lab

Western blot - Anti-Actin antibody [IGX3831H] - Loading Control (AB213251)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab213251 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

All lanes:

Anti-Actin [IGX3831H] at 1 µg/mL

Lane 1:

Human colon tissue lysate at 10 µg

Lane 2:

Mouse colon tissue lysate at 10 µg

Lane 3:

HeLa whole cell lysate at 10 µg

Lane 4:

Jurkat whole cell lysate at 10 µg

Lane 5:

NIH3T3 whole cell lysate at 10 µg

Lane 6:

PANC1 whole cell lysate at 10 µg

Lane 7:

C2C12 whole cell lysate at 10 µg

Lane 8:

Human skeletal muscle tissue lysate at 10 µg

Secondary

All lanes:

HRP conjugated Goat Anti-Human IgG (H+L) at 1/10000 dilution

Predicted band size: 42 kDa

false

Exposure time: 1min