Anti-beta Actin antibody [AC-15] (ab6276)

  Western blot - beta Actin antibody [AC-15] (ab6276)

Predicted band size : 42 kDa


MDCK cells induced with increasing amounts of doxycycline to control expression of the gene of interest. All cells were normalized for loading with an albumin protein standard asssay. Anti-beta actin (ab6276) was used at a concentration of 1:5000 in a milk blocking solution. B-actin blotting confirms the albumin assay in showing that an equal amount of lysate was loaded in each lane.

David Grimm, Yale University, USA.

  Western blot - beta Actin antibody [AC-15] (ab6276)

All lanes : Anti-beta Actin antibody [AC-15] (ab6276) at 1/5000 dilution

Lane 1 : HeLa nuclear
Lane 2 : HeLa whole cell lysate
Lane 3 : A431 cell lysate
Lane 4 : Jurkat cell lysate
Lane 5 : HEK293 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
Alexa Fluor anti mouse at 1/5000 dilution

Performed under reducing conditions.

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

  Immunocytochemistry/ Immunofluorescence - beta Actin antibody [AC-15] (ab6276)

ab6276 at 1/500 staining human macrophage cells by ICC/IF. The cells were paraformaldehyde fixed and blocked with serum prior to incubation with the antibody for 1 hour. A Cy2 ® conjugated donkey antibody was used as the secondary.

This image is courtesy of an anonymous Abreview

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - beta Actin antibody [AC-15] (ab6276)

ab6276 at 1/10000 staining mouse brain tissue sections by IHC-P. The mouse brain was immersion-fixed in 4% Formalin/PBS for 2 days, then bisected mid-saggitally. The tissue was further fixed in same solution for a further 3 days. Both halves were processed to pwax on a dip-n-dunk tissue processor using a 20hr-cycle. Sections were cut at 5microns,floated out on water at 47C, mounted on Superfrost Plus slides and dried/melted for 24hrs in a 62C oven.

The whole brain shows positivity but there are areas/cells that are enriched. Note that the axonal tracks are negatively stained at this dilution factor, yet some cells within those tracks are still strongly positive (many similar cells outside of the tracks are also positive).

This image is courtesy of an Abreview submitted by Mr Carl hobbs

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  Western blot - beta Actin antibody [AC-15] (ab6276)

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

Western Blot of ab6276 (used as loading control) with whole tissue lysate of grey matter from BA20 (temporal cortex).  Ab6276 was diluted 1/50000 and incubated with the sample for 16 hours at 4°C.  5 µg of lysate was loaded onto the gel, which was blocked with 5% milk for 1 hour at 20°C.  An Alexa Fluor® 680 conjugated goat anti-mouse antibody, diluted 1/5000, was used as the secondary.

Bands below actin in image are synaptophysin (SYN).

This image is courtesy of an Abreview submitted by Dr Mark Elliott

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  Immunocytochemistry/ Immunofluorescence - beta Actin antibody [AC-15] (ab6276)

ab6276 staining beat actin in rat hypothalamus primary cells by ICC/IF. The cells were formaldehyde fixed, permeabilized in 0.5% Triton X-100 and blocked in 5% serum for 20 minutes at 25°C. The primary antibody was diluted 1/1000 in PBS (0.1% Triton X-100, 1% goat serum) and incubated with sample for 16 hours at 4°C. An Alexa Fluor® 546 conjugated goat polyclonal to mouse IgG1, diluted 1/500 was used as secondary.

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  ELISA - beta Actin antibody [AC-15] (ab6276)

ab6276 detecting beta actin in AGS Human gastric carcinoma cells by direct ELISA. Samples were blocked with 5% BSA for 1 hour at 23°C and incubated with the primary antibody (1/1000 in blocking buffer) for 16 hours at 4°C. Ab6729 (1/1000) was used as the secondary antibody.

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