Immunocytochemistry/ Immunofluorescence - alpha Tubulin antibody (ab18251)

ICC/IF image of ab18251 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab18251, 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).

Western blot - alpha Tubulin antibody (ab18251)

All lanes : Anti-alpha Tubulin antibody (ab18251) at 0.5 µg/ml

Lane 1 : HeLa lysate
Lane 2 : A431 lysate

Lysates/proteins at 20 µg per lane.

Secondary
Alexa Flour Goat polyclonal to Rabbit IgG (700) at 1/10000 dilution

Predicted band size : 50 kDa
Observed band size : 50 kDa
Additional bands at : 30 kDa (possible cross reactivity).

ab18251 detects a strong band at 50 kDa corresponding to alpha tubulin. Cross-reactivity is also seen with other lower molecular weight bands. This may be reduced by using the antibody at a lower working concentration.

Immunocytochemistry/ Immunofluorescence - alpha Tubulin antibody (ab18251)

ab18251 at a 1/8000 dilution staining human HeLa cells by immunocytochemistry. The cells were paraformaldehyde fixed and incubated with the antibody for 30 minutes. The secondary antibody was a Cy3 conjugated Goat Anti-Rabbit IgG (H+L). The image shows staining of an interphase IM cell.

This image is courtesy of an Abreview by Kirk McManus submitted on 27 February 2006.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - alpha Tubulin antibody (ab18251)

Image courtesy of Human Protein Atlas

ab18251 staining alpha Tubulin. Paraffin embedded human appendix tissue was incubated with ab18251 (1/2500 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab18251 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

Immunocytochemistry/ Immunofluorescence - alpha Tubulin antibody (ab18251)

ab18251 staining alpha Tubulin in HeLa cells by Immunocytochemistry/ Immunofluorescence.

ab18251 staining alpha Tubulin labeled red.
ab20740 staining Lamin A labeled green.
Hoechst 33258 staining DNA blue.

Image from Mackay DR et al, J Cell Biol. 2010 Nov 29;191(5):923-31. Epub 2010 Nov 22, Fig 1. DOI 10.1083/jcb.201007124