Antibodies, Proteins, Kits and Reagents for Life Science

Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19591] to beta III Tubulin - BSA and Azide free
Suitable for: IHC-P, WB, Flow Cyt, ICC/IF, IP
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free
See all beta III Tubulin primary antibodies
Description

Rabbit monoclonal [EPR19591] to beta III Tubulin - BSA and Azide free
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HCT116, HeLa, HEK-293 and HAP1 cell lysates. IHC-P: Human cerebral cortex, glioma, lung adenocarcinoma, and cholangiocarcinoma tissues, mouse cerebral cortex, and rat cerebral cortex tissues. IP: U-87 MG cell lysate. Flow Cyt: U-87 cells. ICC/IF: SH-SY5Y and U-87 cells.
General notes
ab238454 is the carrier-free version of ab215037. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab238454 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C. Do Not Freeze.
Storage buffer

pH: 7.2
Constituent: PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19591
Isotype

IgG
Research areas

Western blot - Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

All lanes : Anti-beta III Tubulin antibody [EPR19591] (ab215037) at 1/2000 dilution

Lane 1 : Wild-type HCT116 cell lysate
Lane 2 : TUBB3 knockout HCT116 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 50 kDa
Observed band size: 52 kDa
why is the actual band size different from the predicted?

This data was developed using the same antibody clone in a different buffer formulation (ab215037).

Lanes 1- 4: Merged signal (red and green). Green - ab215037 observed at 52 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab215037 was shown to react with beta III Tubulin in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266900 (knockout cell lysate ab257070) was used. Wild-type HCT116 and TUBB3 knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab215037 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling beta III Tubulin with ab215037 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Cytoplasmic staining on human cerebral cortex is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215037).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

All lanes : Anti-beta III Tubulin antibody [EPR19591] (ab215037) at 1/2000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : TUBB3 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 50 kDa
Observed band size: 50 kDa

This data was developed using the same antibody clone in a different buffer formulation (ab215037).

Lanes 1- 2: Merged signal (red and green). Green - ab215037 observed at 50 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab215037 was shown to react with beta III tubulin in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255358 (knockout cell lysate ab263857) was used. Wild-type HeLa and TUBB3 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab215037 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

All lanes : Anti-beta III Tubulin antibody [EPR19591] (ab215037) at 1/2000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : TUBB3 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : HEK293 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 50 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab215037 observed at 50 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab215037 was shown to specifically react with TUBB3 in wild-type HAP1 cells as signal was lost in TUBB3 knockout cells. Wild-type and TUBB3 knockout samples were subjected to SDS-PAGE. ab215037 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/2000 dilution and 1/20,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing tissue culture supernatant, tris glycine, BSA, glycerol and sodium azide (ab215037).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling beta III Tubulin with ab215037 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Cytoplasmic staining on human glioma is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215037).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

Immunohistochemical analysis of paraffin-embedded human cholangiocarcinoma tissue labeling beta III Tubulin with ab215037 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Cytoplasmic staining on human cholangiocarcinoma is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215037).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling beta III Tubulin with ab215037 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Negative staining on human hepatocellular carcinoma. (PMID: 25039376).

Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215037).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma tissue labeling beta III Tubulin with ab215037 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Cytoplasmic staining on human lung adenocarcinoma is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215037).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling beta III Tubulin with ab215037 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Cytoplasmic staining on mouse cerebral cortex is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215037).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue labeling beta III Tubulin with ab215037 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Cytoplasmic staining on rat cerebral cortex is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215037).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling beta III Tubulin with ab215037 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining on SH-SY5Y cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215037).
Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling beta III Tubulin with ab215037 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining on U-87 MG cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215037).
Flow Cytometry - Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

Flow cytometric analysis of 4% paraformaldehyde-fixed U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling beta III Tubulin with ab215037 at 1/200 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215037).
Immunoprecipitation - Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

beta III Tubulin was immunoprecipitated from 0.35 mg of U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate with ab215037 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab215037 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: U-87 MG whole cell lysate 10µg (Input).

Lane 2: ab215037 IP in U-87 MG whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab215037 in U-87 MG whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 10 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215037).
Anti-beta III Tubulin antibody [EPR19591] - BSA and Azide free (ab238454)

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