Anti-Tubulin antibody [YL1/2] - Loading Control

• WB

Lab

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

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 5% Milk before being incubated with ab6160 overnight at 4°C. Antibody binding was detected using ab175786 at a 1 : 10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

All lanes:

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/mL

All lanes:

HeLa Whole Cell Lysate at 10 µg/mL

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (Alexa Fluor® 790) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-alexa-fluor-790-ab175786'>ab175786</a>) at 1/10000 dilution

Predicted band size: 36 kDa

Observed band size: 52 kDa

false

• WB

Unknown

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

Lanes 1 and 3:

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1/5000 dilution

Lanes 2 and 4:

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1/10000 dilution

Lanes 1 - 2:

HeLa (Human epithelial carcinoma cell line) whole cell lysate at 20 µg

Lanes 3 - 4:

Western blot - BALB/3T3 whole cell lysate (<a href='/en-us/products/cell-lysates/balb-3t3-whole-cell-lysate-ab7901'>ab7901</a>) at 20 µg

Secondary

All lanes:

Western blot - Rabbit Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/rabbit-rat-igg-h-l-hrp-ab6734'>ab6734</a>) at 1/2000 dilution

Predicted band size: 50 kDa

Observed band size: 17 kDa,34 kDa,52 kDa,80 kDa

false

Exposure time: 10s

• WB

Unknown

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

All lanes:

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/mL

All lanes:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Rabbit Anti-Rat IgG H&L (Alkaline Phosphatase) (<a href='/en-us/products/secondary-antibodies/rabbit-rat-igg-h-l-alkaline-phosphatase-ab102169'>ab102169</a>) at 1/2000 dilution

Lanes 3 - 4:

Western blot - Rabbit Anti-Rat IgG H&L (Alkaline Phosphatase) (<a href='/en-us/products/secondary-antibodies/rabbit-rat-igg-h-l-alkaline-phosphatase-ab102169'>ab102169</a>) at 1/10000 dilution

Lanes 5 - 6:

Western blot - Rabbit Anti-Rat IgG H&L (Alkaline Phosphatase) (<a href='/en-us/products/secondary-antibodies/rabbit-rat-igg-h-l-alkaline-phosphatase-ab102169'>ab102169</a>) at 1/20000 dilution

Predicted band size: 36 kDa

Observed band size: 52 kDa

true

• WB

Lab

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

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 5% Milk before being incubated with ab6160 overnight at 4°C. Antibody binding was detected using ab175778 at a 1 : 10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

All lanes:

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/mL

All lanes:

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

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (Alexa Fluor® 680) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-alexa-fluor-680-ab175778'>ab175778</a>) at 1/10000 dilution

Predicted band size: 36 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

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 5% Milk before being incubated with ab6160 overnight at 4°C. Antibody binding was detected using ab175777 at a 1 : 10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

All lanes:

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/mL

Lane 1:

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

Lane 2:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 5 µg

Lane 3:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 2 µg

Lane 4:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 1 µg

Secondary

All lanes:

Western blot - Donkey Anti-Rat IgG H&L (Alexa Fluor® 680) preadsorbed (<a href='/en-us/products/secondary-antibodies/donkey-rat-igg-h-l-alexa-fluor-680-preadsorbed-ab175777'>ab175777</a>) at 1/10000 dilution

Predicted band size: 36 kDa

Observed band size: 52 kDa

false

• WB

Lab

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

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 5% Milk before being incubated with ab6160 overnight at 4°C. Antibody binding was detected using ab175751 at a 1 : 10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

All lanes:

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/mL

All lanes:

HeLa Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (Alexa Fluor® 750) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-alexa-fluor-750-ab175751'>ab175751</a>) at 1/10000 dilution

Predicted band size: 36 kDa

Observed band size: 50 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

ICC/IF image of ab6160 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

The cells were fixed in 100% methanol 5 minutes, permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6160, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was ab150165 Alexa Fluor^® 488 goat anti-rat IgG (H+L) pre-adsorbed, used at a 1/1000 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10 minutes).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with ab6160 (red line).

The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 (ab6160, 1 μg/1x10⁶ cells) for 30 minutes at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was rat IgG2a [aRTK2758] (ab18450, 1 μg/1x10⁶ cells) used under the same conditions.

Acquisition of >5,000 events was performed.

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

Cytoskeleton and major extracellular matrix proteins in human DPSC (Dental pulp stem cell) were analyzed by immunofluorescence.

Vimentin and tubulin (Panel D, control, E, (BD) and F, (BR)) are shown.

After 7 days in contact with/without the materials (Biodentine (BD) and Bioroot (BR)), coated coverslip cultures were fixed in PBS (pH 7.4) containing 4% paraformaldehyde/5% sucrose for 10 minutes. For detection of intracellular molecules, the cells on the coverslips were permeabilized using 0.5% Triton X-100. To block background staining, cells were treated with PBS containing 1% BSA/1% glycine at 37°C for 20 minutes. Samples were incubated with the primary antibody at 4°C overnight or at 37°C for 2 hours. For double immunostaining, primary antibodies were incubated as above. Samples were then incubated with the appropriate secondary antibodies at 37°C for 1 hour. Cell nuclei were stained using DAPI.

Scale Bar : 100 μm.

Loison-Robert et al PLoS One. 2018 Jan 25;13(1):e0190014. doi: 10.1371/journal.pone.0190014. eCollection 2018. Fig 5. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

IHC image of Tubulin staining in human colon formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer pH 6 for 20 minutes. The section was then incubated with ab6160, 5 μg/ml, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• WB

Lab

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

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 5% milk blocking buffer before being incubated with ab6160 overnight at 4°C. Antibody binding was detected using ab175750 at a 1 : 10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

All lanes:

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/mL

All lanes:

HeLa Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Donkey Anti-Rat IgG H&L (Alexa Fluor® 750) preadsorbed (<a href='/en-us/products/secondary-antibodies/donkey-rat-igg-h-l-alexa-fluor-750-preadsorbed-ab175750'>ab175750</a>) at 1/10000 dilution

Predicted band size: 36 kDa

false

• WB

Unknown

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

Additional bands at : 85 kDa. We are unsure as to the identity of these extra bands.

All lanes:

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/mL

All lanes:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab97057'>ab97057</a>) at 1/2000 dilution

Lanes 3 - 4:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab97057'>ab97057</a>) at 1/10000 dilution

Lanes 5 - 6:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab97057'>ab97057</a>) at 1/20000 dilution

Predicted band size: 36 kDa

Observed band size: 52 kDa

true

• WB

Unknown

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

All lanes:

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/mL

All lanes:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Rabbit Anti-Rat IgG H&L (Alkaline Phosphatase) (<a href='/en-us/products/secondary-antibodies/rabbit-rat-igg-h-l-alkaline-phosphatase-ab6735'>ab6735</a>) at 1/2000 dilution

Lanes 3 - 4:

Western blot - Rabbit Anti-Rat IgG H&L (Alkaline Phosphatase) (<a href='/en-us/products/secondary-antibodies/rabbit-rat-igg-h-l-alkaline-phosphatase-ab6735'>ab6735</a>) at 1/10000 dilution

Lanes 5 - 6:

Western blot - Rabbit Anti-Rat IgG H&L (Alkaline Phosphatase) (<a href='/en-us/products/secondary-antibodies/rabbit-rat-igg-h-l-alkaline-phosphatase-ab6735'>ab6735</a>) at 1/20000 dilution

Predicted band size: 36 kDa

true

• WB

Unknown

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

All lanes:

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/mL

All lanes:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Goat Anti-Rat IgG H&L (Alkaline Phosphatase) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-alkaline-phosphatase-ab6846'>ab6846</a>) at 1/5000 dilution

Lanes 3 - 4:

Western blot - Goat Anti-Rat IgG H&L (Alkaline Phosphatase) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-alkaline-phosphatase-ab6846'>ab6846</a>) at 1/20000 dilution

Lanes 5 - 6:

Western blot - Goat Anti-Rat IgG H&L (Alkaline Phosphatase) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-alkaline-phosphatase-ab6846'>ab6846</a>) at 1/50000 dilution

Predicted band size: 36 kDa

true

• WB

Unknown

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

Lanes 1 - 6:

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/mL

Lane 5:

Anti-RUNX2 antibody (<a href='/en-us/products/unavailable/runx2-antibody-ab102712'>ab102712</a>) at 1/20000 dilution

All lanes:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Rabbit Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/rabbit-rat-igg-h-l-hrp-ab102172'>ab102172</a>) at 1/2000 dilution

Lanes 3 - 4:

Western blot - Rabbit Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/rabbit-rat-igg-h-l-hrp-ab102172'>ab102172</a>) at 1/10000 dilution

Lane 6:

Western blot - Rabbit Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/rabbit-rat-igg-h-l-hrp-ab102172'>ab102172</a>) at 1/20000 dilution

Predicted band size: 36 kDa

Observed band size: 52 kDa

true

• WB

Project9436****

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (AB6160)

All lanes:

Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) whole cell lysate at 10 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

Lane 3:

PC-12 (Rat adrenal pheochromocytoma cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Peroxidase Conjugated Rabbit Anti-Rat IgG (H+L) at 1/10000 dilution

Predicted band size: 50 kDa

Observed band size: 52 kDa,85 kDa

false

Exposure time: 8min