Anti-Tubulin 抗体 [YOL1/34] - Microtubule Marker

• WB

Lab

Western blot - Anti-Tubulin antibody [YOL1/34] - Loading Control (AB6161)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab6161 overnight at 4°C. Antibody binding was detected using ab205720, and visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-Tubulin antibody [YOL1/34] - Loading Control (ab6161) at 1 µg/mL

Lane 1:

Liver (Human) Tissue Lysate at 10 µg

Lane 2:

Liver (Mouse) Tissue Lysate at 10 µg

Lane 3:

Liver (Rat) Tissue Lysate at 10 µg

Lane 4:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 5:

NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Lane 6:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 36 kDa

Observed band size: 54 kDa

true

Exposure time: 15s

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Tubulin antibody [YOL1/34] - Loading Control (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 22°C. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22°C. 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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Tubulin antibody [YOL1/34] - Loading Control (AB6161)

ab6161 staining Tubulin in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab6161 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150165, Goat polyclonal Secondary Antibody to Rat IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Tubulin antibody [YOL1/34] - Loading Control (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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Tubulin antibody [YOL1/34] - Loading Control (AB6161)

ab6161 staining tubulin HeLa cells treated with anisomycin (ab120495), by ICC/IF. Increase in tubulin expression correlates with increased concentration of anisomycin as described in literature.
The cells were incubated at 37°C for 6h in media containing different concentrations of ab120495 (anisomycin) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab6161 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rat polyclonal antibody (ab98386) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

• ICC/IF

AbReview11376****

Immunocytochemistry/ Immunofluorescence - Anti-Tubulin antibody [YOL1/34] - Loading Control (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

• WB

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Western blot - Anti-Tubulin antibody [YOL1/34] - Loading Control (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).

All lanes:

Western blot - Anti-Tubulin antibody [YOL1/34] - Loading Control (ab6161)

Predicted band size: 50 kDa

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• WB

Lab

Western blot - Anti-Tubulin antibody [YOL1/34] - Loading Control (AB6161)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab6161 overnight at 4°C. Antibody binding was detected using ab205720 (Left Image) and a competitor secondary (Right Image), and visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-Tubulin antibody [YOL1/34] - Loading Control (ab6161) at 1 µg/mL

Lane 1:

Liver (Human) Tissue Lysate at 10 µg

Lane 2:

Liver (Mouse) Tissue Lysate at 10 µg

Lane 3:

Liver (Rat) Tissue Lysate at 10 µg

Secondary

All lanes:

<a href='/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a> (Left Image) at 1/5000 and a competitor secondary (Right Image) at 1/5000. Notice the increased background of the competitor product

Predicted band size: 36 kDa

Observed band size: 54 kDa

false

Exposure time: 15s

• WB

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Western blot - Anti-Tubulin antibody [YOL1/34] - Loading Control (AB6161)

All lanes:

Western blot - Anti-Tubulin antibody [YOL1/34] - Loading Control (ab6161) at 1 µg/mL

All lanes:

Brain (Rat) Tissue Lysate at 10 µg

Secondary

All lanes:

Rabbit polyclonal to Rat IgG - H&L (HRP) at 1/10000 dilution

Predicted band size: 50 kDa,55 kDa

Observed band size: 54 kDa

true

Exposure time: 3min

• WB

AbReview7034****

Western blot - Anti-Tubulin antibody [YOL1/34] - Loading Control (AB6161)

All lanes:

Western blot - Anti-Tubulin antibody [YOL1/34] - Loading Control (ab6161) at 1/2000 dilution

All lanes:

Yeast (Saccharomyces cerevisiae) whole cell extract prepared by bead-beating at 5 µg

Secondary

All lanes:

HRP conjugated goat anti-rat antibody

Predicted band size: 50 kDa

Observed band size: 50 kDa

true

Exposure time: 30s

This image is courtesy of an anonymous Abreview