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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: 20
Lane 2: 20
Lane 3: 20
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.
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.
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
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.
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
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
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.
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) (
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