- Tubulin antibody [YOL1/34] - Microtubule Marker (ab6161)

Western blot against tubulin with ab6161 at 1/3000.  Secondary Rabbit anti-Rat IgG HRP (ab6734)was used at 1/2000.  Exposure time: 2mins.

Lane 1: 20µg/lane HeLa (Human) whole cell lysates (ab7898).

Lane 2: 20µg/lane 3T3 (Mouse) whole cell lysate (ab7901).

Lane 3: 20µg/lane Rat brain tissue lysate (ab7942).

- Tubulin antibody [YOL1/34] - Microtubule Marker (ab6161)

Cultured human macrophages were used with ab6161 at 1/1000 for immunofluorescence. Cells were fixed with cold 2% formaldehyde for 20mins.

Green staining is Alexa 568, Blue staining is DAPI stain.

This cell represents a young macrophage, the staining patterns varied as the cells aged in culture.

- Tubulin antibody [YOL1/34] - Microtubule Marker (ab6161)

Confocal image of  21 day in vitro rat hippocampal neurons, stained with rat monoclonal antibody to Tubulin - Microtubule Marker (ab6161) in green at 1/500 and Microtubule Associated protein 2 in blue.

This picture was kindly supplied as part of the review submitted by Dr Jonathon Burman.

Western blot - Tubulin antibody [YOL1/34] - Microtubule Marker (ab6161)

All lanes : Anti-Tubulin antibody [YOL1/34] - Microtubule Marker (ab6161) at 1/2000 dilution

Lane 1 : Yeast whole cell extract prepared by bead-beating
Lane 2 : Yeast whole cell extract prepared by bead-beating
Lane 3 : Yeast whole cell extract prepared by bead-beating
Lane 4 : Yeast whole cell extract prepared by bead-beating
Lane 5 : Yeast whole cell extract prepared by bead-beating
Lane 6 : Yeast whole cell extract prepared by bead-beating
Lane 7 : Yeast whole cell extract prepared by bead-beating
Lane 8 : Yeast whole cell extract prepared by bead-beating
Lane 9 : Yeast whole cell extract prepared by bead-beating
Lane 10 : Yeast whole cell extract prepared by bead-beating

Lysates/proteins at 5 µg per lane.

Secondary
HRP conjugated goat anti-rat antibody
developed using the ECL technique

Performed under reducing conditions.

Observed band size : 50 kDa (why is the actual band size different from the predicted?)


Exposure time : 30 seconds

This image is courtesy of an anonymous Abreview

Immunocytochemistry/ Immunofluorescence - Tubulin antibody [YOL1/34] - Microtubule Marker (ab6161)

ICC/IF image of ab6161 stained human HepG2 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab6161, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rat IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, HEK 293 and MCF7 cells.

Immunocytochemistry/ Immunofluorescence - Tubulin antibody [YOL1/34] - Microtubule Marker (ab6161)

ab6161 staining mouse NIH 3T3 fibroblast cells by ICC/IF. Cells were PFA fixed  and permeabilized in 0.2% Triton X-100 prior to blocking in 5% BSA for 45 minutes at RT.  The primary antibody was diluted 1/1000 and incubated with the sample for 1 hour.  An Alexa Fluor® 568 conjugated goat anti-rat antibody, diluted 1/3000, was used as the secondary.

This image is courtesy of an anonymous Abreview

Western blot - Tubulin antibody [YOL1/34] - Microtubule Marker (ab6161)

Anti-Tubulin antibody [YOL1/34] - Microtubule Marker (ab6161) at 1 µg/ml + Brain (Rat) Tissue Lysate at 10 µg

Secondary
Rabbit polyclonal to Rat IgG - H&L (HRP) at 1/10000 dilution
developed using the ECL technique

Performed under reducing conditions.

Observed band size : 54 kDa (why is the actual band size different from the predicted?)


Exposure time : 3 minutes

Immunocytochemistry/ Immunofluorescence - Tubulin antibody [YOL1/34] - Microtubule Marker (ab6161)

ICC/IF image of ab6161 stained Hela cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6161, 1µg/ml) overnight at +4ºC. The secondary antibody (green) was DyLight® 488 goat anti-rat- H&L, pre-adsorbed (ab98420) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Flow Cytometry-Anti-Tubulin antibody [YOL1/34] - Microtubule Marker(ab6161)

Overlay histogram showing HeLa cells stained with ab6161 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab6161, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG2a [aRTK2758] (ab18450, 1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.