Western blot - alpha Tubulin antibody - Loading Control (ab4074)

All lanes : Anti-alpha Tubulin antibody - Loading Control (ab4074) at 1 µg/ml

Lane 1 : Mouse 3T3 lysate
Lane 2 : Rat liver lysate
Lane 3 : Cow cell lysate
Lane 4 : Chicken cell lysate
Lane 5 : CHO lysate

Lysates/proteins at 20 µg per lane.

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 50 kDa
Observed band size : 50 kDa


Exposure time : 30 seconds

ab4074 cross-reacts strongly with Mouse 3T3 lysate and shows weak cross-reactivity with Rat liver, Cow, Chicken and CHO lysates.

Immunocytochemistry/ Immunofluorescence - alpha Tubulin antibody - Loading Control (ab4074)

ICC/IF image of ab4074 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab4074, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT^TM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 phalloidin was used to label F-actin (red). 

Western blot - alpha Tubulin antibody - Loading Control (ab4074)

All lanes : Anti-alpha Tubulin antibody - Loading Control (ab4074) at 1 µg/ml

Lane 1 : HeLa whole cell lysate
Lane 2 : HeLa whole cell lysate with alpha Tubulin peptide (ab23537) at 1 µg/ml

Lysates/proteins at 20 µg per lane.

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution

Predicted band size : 50 kDa
Observed band size : 50 kDa

Immunohistochemistry (Paraffin-embedded sections) - alpha Tubulin antibody - Loading Control (ab4074)

(Image courtesy of Human Protein Atlas).
ab4074 staining alpha tubulin in the glandular cells of the endometrium. Paraffin embedded endometrium tissue of a 33 year-old female was incubated with ab4074 (1/25 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

ab4074 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org

Western blot - alpha Tubulin antibody - Loading Control (ab4074)

Anti-alpha Tubulin antibody - Loading Control (ab4074) at 1 µg/ml + whole cell lysate prepared from MCF7 cells at 20 µg

Secondary
Goat anti-rabbit IgG-HRP at 1/1000 dilution
developed using the ECL technique

Predicted band size : 50 kDa
Observed band size : 50 kDa


Exposure time : 5 minutes

Image courtesy of an anonymous Abreview.

Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Loading Control (ab4074)

ab4074 staining alpha Tubulin in murine sperm by Immunocytochemistry/ Immunofluorescence.

Cells were fixed in 2% paraformaldehyde, permeabilized in -20°C methanol for 10 minutes, blocked using 3% BSA for 1 hour at 25°C and then incubated with ab4074 at a 1/300 dilution for 2 hours at 25°C. The secondary used was an Alexa-Fluor 546 conjugated goat anti-rabbit (H+L) polyclonal used at a1/400 dilution.

Image courtesy of an anonymous Abreview.