Anti-Tubulin (acetyl K40) antibody [6-11B-1] (ab11323)

  Immunocytochemistry/ Immunofluorescence - Tubulin (acetyl K40) antibody [6-11B-1] (ab11323)

ab11323 at 1/800 staining dog kidney cells by ICC/IF. The cells were paraformaldehyde fixed and blocked with BSA prior to incubation with the antibody for 1 hour. An Alexa-Fluor ® 488 conjugated goat anti-mouse IgG was used as the secondary.

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  Western blot - Tubulin (acetyl K40) antibody [6-11B-1] (ab11323)

All lanes : Anti-Tubulin (acetyl K40) antibody [6-11B-1] (ab11323) at 1/1000 dilution

Lane 1 : Whole animal lysate prepared from wild type (N2) C. Elegans worms.
Lane 2 : Whole animal lysate prepared from elpc-1 C. Elegans worms.
Lane 3 : Whole animal lysate prepared from elpc-3 C. Elegans worms.
Lane 4 : Whole animal lysate prepared from mec-12 (e1605) C. Elegans worms.
Lane 5 : Whole animal lysate prepared from mec-12 (e1607) C. Elegans worms.

Lysates/proteins at 20 µg per lane.


To measure acetylated α-tubulin levels by western blot, protein was extracted from young adult worms. To avoid protein degradation, worms were suspended in ice-cold extraction buffer containing proteinase inhibitors and rapidly frozen in liquid nitrogen. The frozen pellets were ground to a powder in a mortar.

Image from Chen C et al, PLoS Genet. 2009 Jul;5(7):e1000561. Epub 2009 Jul 10, fig S9.

  Western blot - Tubulin (acetyl K40) antibody [6-11B-1] (ab11323)

Anti-Tubulin (acetyl K40) antibody [6-11B-1] (ab11323) at 1/2000 dilution (in PBST + 2.5% milk for 16 hours at 4°C) + Whole cell lysate of human IHP1 cells at 30 µg

Secondary
An HRP-conjugated Goat anti-mouse IgG monoclonal at 1/2000 dilution
developed using the ECL technique

Performed under reducing conditions.

Observed band size : 55 kDa (why is the actual band size different from the predicted?)


Exposure time : 20 seconds

Blocking Step: 5% Milk for 2 hours at 22°C

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  Immunocytochemistry/ Immunofluorescence - Tubulin (acetyl K40) antibody [6-11B-1] (ab11323)

ab11323 staining Tubulin (acetyl K40) in human glioblastoma tumor sphere cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 0.1% in PBS and blocked with 0.5% BSA for 20 minutes at room temperature. Samples were incubated with primary antibody (1/100 in PBS + 0.5% BSA) for 1 hour at 4°C. An Alexa Fluor^®488-conjugated Goat anti-mouse IgG polyclonal (1/400) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).

This image is courtesy of an abreview submitted by Lars Ruether

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