All lanes : Anti-beta Actin antibody [SP124] (ab115777) at 1/200 dilution

Lane 1 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 4 : A549 (Human lung carcinoma epithelial cell) whole cell lysate
Lane 5 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate
Lane 6 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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


Exposure time: 10 seconds

Blocking/Diluting buffer and concentration 5% NFDM/TBST

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat placenta tissue sections labeling beta Actin with ab115777 at 1/100 dilution. Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with citrate sodium buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse placenta tissue sections labeling beta Actin with ab115777 at 1/200 dilution. Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with citrate sodium buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human placenta tissue sections labeling beta Actin with ab115777 at 1/200 dilution. Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with citrate sodium buffer (pH 6.0, epitope retrieval solution 1) for 10mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Immunocytochemistry/ Immunofluorescence analysis of C6 (rat glial tumor glial cell) cells labeling beta Actin with purified ab115777 at 1/50 dilutioin. Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Flow Cytometry analysis of C6 (rat glial tumor glial cell) cells labeling beta Actin with purified ab115777 at 1/200 - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) was used as the secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue.

Staining of Human beta Actin in a Formalin/PFA-fixed paraffin-embedded section of Human Placenta using ab115777 at a dilution of 1/200.

Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling beta Actin with purified ab115777 at 1/50 dilution. Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Flow Cytometry analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling beta Actin with purified ab115777 at 1/200 - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) was used secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue.

Flow Cytometry analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling beta Actin with purified ab115777 at 1/200 - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081)  was used as the secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue.

Anti-beta Actin antibody [SP124] (ab115777) at 1/200 dilution + Raji cell lystate

Predicted band size: 42 kDa

Flow cytometric analysis of rabbit anti-beta Actin (SP124) antibody ab115777 (1/200) in HeLa cells (green) compared to negative control of rabbit IgG (blue)