Anti-beta II Tubulin antibody [EPR16773] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-beta II Tubulin antibody [EPR16773] - BSA and Azide free (AB250105)

This data was developed using ab179512, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed SH-SY5Y cells labeling beta II Tubulin with ab179512 at 1/450 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta II Tubulin antibody [EPR16773] - BSA and Azide free (AB250105)

This data was developed using ab179512, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling beta II Tubulin with ab179512 at 1/250 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. High level of cytoplasmic and nuclear expression on neuron cells of Human cerebral cortex is observed. Counter stained with Hematoxylin.

Negative control : PBS instead of primary antibody; secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-beta II Tubulin antibody [EPR16773] - BSA and Azide free (AB250105)

This data was developed using ab179512, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling beta II Tubulin with ab179512 at 1/100 dilution, followed by anti-rabbit Alexa Fluor® 488 (ab150077) secondary antibody at 1/400 dilution (green). Confocal image shows both cytoplasm and nuclear staining on the SH-SY5Y cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and anti-mouse AlexaFluor® 594 (ab150120) at 1/500 dilution (red).

The negative controls are as follows : -

-ve control 1 : - ab179512 at 1/100 dilution followed by anti-mouse AlexaFluor® 594 (ab150120) at 1/500 dilution.

-ve control 2 : - ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by anti-rabbit Alexa Fluor® 488 (ab150077) at 1/400 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta II Tubulin antibody [EPR16773] - BSA and Azide free (AB250105)

This data was developed using ab179512, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human glioma labeling beta II Tubulin with ab179512 at 1/250 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasmic staining on Human glioma cells is observed. Counter stained with Hematoxylin.

Negative control : PBS instead of primary antibody; secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-beta II Tubulin antibody [EPR16773] - BSA and Azide free (AB250105)

This data was developed using ab179512, the same antibody clone in a different buffer formulation.

Beta II Tubulin was immunoprecipitated from 1mg of Human fetal brain extract with ab179512 at 1/140 dilution. Western blot was performed from the immunoprecipitate using ab179512 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : Human fetal brain extract. Lane 2 : PBS instead of extract.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-beta II Tubulin antibody [EPR16773] (<a href='/en-us/products/primary-antibodies/beta-ii-tubulin-antibody-epr16773-ab179512'>ab179512</a>)

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta II Tubulin antibody [EPR16773] - BSA and Azide free (AB250105)

This data was developed using ab179512, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling beta II Tubulin with ab179512 at 1/250 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. High level of expression on neuron cells of Rat cerebral cortex is observed. Counter stained with Hematoxylin.

Negative control : PBS instead of primary antibody; secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta II Tubulin antibody [EPR16773] - BSA and Azide free (AB250105)

This data was developed using ab179512, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling beta II Tubulin with ab179512 at 1/250 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. High level of expression on neuron cells of Mouse cerebral cortex is observed. Counter stained with Hematoxylin.

Negative control : PBS instead of primary antibody; secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-beta II Tubulin antibody [EPR16773] - BSA and Azide free (AB250105)

This data was developed using ab179512, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Beta II Tubulin is highly expressed in brain tissues and cell lines derived from the central nervous system.

All lanes:

Western blot - Anti-beta II Tubulin antibody [EPR16773] (<a href='/en-us/products/primary-antibodies/beta-ii-tubulin-antibody-epr16773-ab179512'>ab179512</a>) at 1/1000 dilution

Lane 1:

Human fetal heart lysate at 10 µg

Lane 2:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-beta II Tubulin antibody [EPR16773] - BSA and Azide free (AB250105)

False colour image of Western blot : Anti-beta II Tubulin antibody [EPR16773] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab179512 was shown to bind specifically to beta II Tubulin. A band was observed at 50 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in Tubb2b knockout cell line ab266692 (knockout cell lysate ab258741). To generate this image, wild-type and Tubb2b knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

Lanes 1 - 2:

Western blot - Anti-beta II Tubulin antibody [EPR16773] (<a href='/en-us/products/primary-antibodies/beta-ii-tubulin-antibody-epr16773-ab179512'>ab179512</a>) at 1/1000 dilution

Lanes 3 - 4:

Western blot - Anti-beta II Tubulin antibody [EPR16773] - BSA and Azide free (ab250105) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

TUBB2A knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human TUBB2A knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-tubb2a-knockout-hek-293t-cell-line-ab266692'>ab266692</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

THP-1 cell lysate at 20 µg

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

• WB

Supplier Data

Western blot - Anti-beta II Tubulin antibody [EPR16773] - BSA and Azide free (AB250105)

This data was developed using ab179512, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Beta II Tubulin is highly expressed in brain tissues and cell lines derived from the central nervous system.

All lanes:

Western blot - Anti-beta II Tubulin antibody [EPR16773] (<a href='/en-us/products/primary-antibodies/beta-ii-tubulin-antibody-epr16773-ab179512'>ab179512</a>) at 1/10000 dilution

All lanes:

Human fetal brain lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

• WB

Supplier Data

Western blot - Anti-beta II Tubulin antibody [EPR16773] - BSA and Azide free (AB250105)

This data was developed using ab179512, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Beta II Tubulin is highly expressed in brain tissues and cell lines derived from the central nervous system.

All lanes:

Western blot - Anti-beta II Tubulin antibody [EPR16773] (<a href='/en-us/products/primary-antibodies/beta-ii-tubulin-antibody-epr16773-ab179512'>ab179512</a>) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse heart lysate at 10 µg

Lane 3:

Mouse kidney lysate at 10 µg

Lane 4:

Mouse spleen lysate at 10 µg

Lane 5:

Rat brain lysate at 10 µg

Lane 6:

Rat heart lysate at 10 µg

Lane 7:

Rat kidney lysate at 10 µg

Lane 8:

C6 (Rat glial tumor cells) whole cell lysate at 10 µg

Lane 9:

PC12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 50 kDa

Observed band size: 50 kDa

false