Name: Anti-beta Tubulin antibody [EPR16774] - Loading Control
SKU: ab179513
Rating: 5 (2 reviews)

Rabbit Recombinant Monoclonal Beta-5-tubulin antibody. Suitable for WB, Flow Cyt (Intra), IHC-P, ICC/IF and reacts with Recombinant full length protein - Human, Rat, Mouse, African green monkey, Drosophila melanogaster, Zebrafish, Xenopus tropicalis, Chicken, Human, Dog, Cow samples. Cited in 79 publications.

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Images

Western blot - Anti-beta Tubulin antibody [EPR16774] - Loading Control (AB179513), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	Flow Cyt (Intra)	IHC-P	ICC/IF
Human	Tested	Tested	Tested	Tested
Mouse	Tested	Expected	Tested	Tested
Rat	Tested	Expected	Tested	Expected
African green monkey	Tested	Expected	Expected	Expected
Chicken	Tested	Expected	Expected	Expected
Cow	Tested	Expected	Expected	Expected
Dog	Tested	Expected	Expected	Expected
Drosophila melanogaster	Tested	Expected	Expected	Expected
Monkey	Predicted	Predicted	Predicted	Predicted
Recombinant full length protein - Human	Tested	Not recommended	Not recommended	Not recommended
Xenopus tropicalis	Tested	Expected	Expected	Expected
Zebrafish	Tested	Expected	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Recombinant full length protein - Human	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -
Species African green monkey	Dilution info 1/1000	Notes -
Species Drosophila melanogaster	Dilution info 1/1000	Notes -
Species Zebrafish	Dilution info 1/1000	Notes -
Species Xenopus tropicalis	Dilution info 1/1000	Notes -
Species Chicken	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -
Species Dog	Dilution info 1/1000	Notes -
Species Cow	Dilution info 1/1000	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Monkey	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/150	Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Rat, Mouse, African green monkey, Drosophila melanogaster, Zebrafish, Xenopus tropicalis, Chicken, Dog, Cow	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Monkey	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant full length protein - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info 1/250	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/250	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/250	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species African green monkey, Drosophila melanogaster, Zebrafish, Xenopus tropicalis, Chicken, Dog, Cow	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Monkey	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant full length protein - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat, African green monkey, Drosophila melanogaster, Zebrafish, Xenopus tropicalis, Chicken, Dog, Cow	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Monkey	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant full length protein - Human	Dilution info -	Notes -

Associated Products

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Target data

See full target information TUBB

Function

Tubulin is the major constituent of microtubules, a cylinder consisting of laterally associated linear protofilaments composed of alpha- and beta-tubulin heterodimers. Microtubules grow by the addition of GTP-tubulin dimers to the microtubule end, where a stabilizing cap forms. Below the cap, tubulin dimers are in GDP-bound state, owing to GTPase activity of alpha-tubulin.

Alternative names

TUBB5, OK/SW-cl.56, TUBB, Tubulin beta chain, Tubulin beta-5 chain

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR16774
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Beta tubulin also known as beta-tubulin or β-tubulin is a globular protein with a molecular weight of approximately 50 kDa. It is a major component of microtubules which are cylindrical structures vital for maintaining cell shape and enabling intracellular transport. This protein is expressed widely in eukaryotic cells and plays a role in mitosis and meiosis by forming the spindle apparatus important for chromosome separation during cell division. The size of beta tubulin allows it to effectively polymerize with alpha tubulin forming heterodimers which are the building blocks of the microtubule network.

Biological function summary

This protein contributes significantly to the dynamic instability of microtubules allowing rapid assembly and disassembly which is essential for cellular processes like motility signaling and maintaining the cell’s architecture. Beta tubulin operates as part of the tubulin family which includes several related proteins within microtubule structures. The beta-tubulin molecules in microtubules are critical for the interactions with microtubule-associated proteins (MAPs) and motor proteins such as kinesin and dynein influencing trafficking and positioning of organelles within the cell.

Pathways

Beta tubulin plays key roles in the mitotic spindle assembly checkpoint ensuring accurate chromosome segregation. It is actively involved in the microtubule pathway and has associations with signaling pathways such as the Wnt signaling pathway which affects cell growth and differentiation. Through these pathways beta tubulin interacts with proteins like tau and MAP2 which stabilize microtubules to control their functional dynamics within the cell.

Associated diseases and disorders

Mutations or dysregulations in beta tubulin can affect neurological and proliferative conditions such as neurodegenerative diseases and cancer. Beta tubulin is notably linked to Alzheimer's disease where tau protein tangles disrupt normal microtubule function. It also connects to diseases like paclitaxel-resistant cancer where altered beta tubulin isoform expression can lead to chemotherapy resistance. The interaction of beta tubulin with tau and alpha tubulin provides further insight into etiological mechanisms of these conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

20 product images

Western blot - Anti-beta Tubulin antibody [EPR16774] - Loading Control (ab179513)
Beta Tubulin Western blot staining of Human beta I Tubulin recombinant protein using rabbit Anti-beta Tubulin antibody
Blocking/Dilution buffer: 5% NFDM/TBST.
The immunogen of ab179513 has 67% identities with beta I Tubulin. The WB image shows it cross-reacts with beta I Tubulin.
Human beta I Tubulin is an in house recombinant protein (aa1-451) with a proprietary tag.
All lanes: Western blot - Anti-beta Tubulin antibody [EPR16774] - Loading Control (ab179513) at 1/1000 dilution
All lanes: Human beta I Tubulin recombinant protein at 0.01 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 49 kDa
Observed band size: 50 kDa
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody [EPR16774] (ab179513)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling beta Tubulin with ab179513 at 1/1000 dilution, followed by anti-rabbit Alexa Fluor® 488 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and anti-mouse AlexaFluor® 594 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/500 dilution (red).
The negative controls are as follows;
-ve control 1: ab179513 at 1/1000 dilution followed by anti-mouse AlexaFluor® 594 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/500 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by anti-rabbit Alexa Fluor® 488 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/500 dilution.
Flow Cytometry (Intracellular) - Anti-beta Tubulin antibody [EPR16774] (ab179513)
Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling beta Tubulin with ab179513 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a 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.
Western blot - Anti-beta Tubulin antibody [EPR16774] - Loading Control (ab179513)
Beta Tubulin Western blot staining using rabbit Anti-beta Tubulin antibody
Blocking/Dilution buffer: 5% NFDM/TBST.
The amino acid sequence of ab179513 immunogen is identical to those of tubulin beta 2A, 2B, 3, 4A, 4B, 5, 6 and 8 at the same region.
All lanes: Western blot - Anti-beta Tubulin antibody [EPR16774] - Loading Control (ab179513) at 1/2000 dilution
Lane 1: Zebrafish whole lysates at 20 µg
Lane 2: Xenopus tropicalis lysates at 20 µg
Lane 3: Drosophila whole lysates at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 49 kDa
Observed band size: 50 kDa
Western blot - Anti-beta Tubulin antibody [EPR16774] - Loading Control (ab179513)
Beta Tubulin Western blot staining using rabbit Anti-beta Tubulin antibody
Blocking/Dilution buffer: 5% NFDM/TBST.
The amino acid sequence of ab179513 immunogen is identical to those of tubulin beta 2A, 2B, 3, 4A, 4B, 5, 6 and 8 at the same region.
All lanes: Western blot - Anti-beta Tubulin antibody [EPR16774] - Loading Control (ab179513) at 1/2000 dilution
Lane 1: UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) whole cell lysates at 20 µg
Lane 2: MDCK (Canine kidney cell line) whole cell lysates at 20 µg
Lane 3: MDBK (Bovine kidney cell line) whole cell lysates at 20 µg
Lane 4: COS-1 (African green monkey kidney fibroblast-like cell line) whole cell lysates at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 49 kDa
Observed band size: 50 kDa
Western blot - Anti-beta Tubulin antibody [EPR16774] - Loading Control (ab179513)
Beta Tubulin Western blot staining using rabbit Anti-beta Tubulin antibody
Blocking/Dilution buffer: 5% NFDM/TBST.
The amino acid sequence of ab179513 immunogen is identical to those of tubulin beta 2A, 2B, 3, 4A, 4B, 5, 6 and 8 at the same region.
All lanes: Western blot - Anti-beta Tubulin antibody [EPR16774] - Loading Control (ab179513) at 1/2000 dilution
Lane 1: HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates at 20 µg
Lane 2: Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates at 20 µg
Lane 3: A431 (Human epidermoid carcinoma) whole cell lysates at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 49 kDa
Observed band size: 50 kDa
Western blot - Anti-beta Tubulin antibody [EPR16774] - Loading Control (ab179513)
Beta Tubulin Western blot staining using rabbit Anti-beta Tubulin antibody
Blocking/Dilution buffer: 5% NFDM/TBST.
The amino acid sequence of ab179513 immunogen is identical to those of tubulin beta 2A, 2B, 3, 4A, 4B, 5, 6 and 8 at the same region.
All lanes: Western blot - Anti-beta Tubulin antibody [EPR16774] - Loading Control (ab179513) at 1/2000 dilution
Lane 1: Human fetal brain lysates at 10 µg
Lane 2: Human fetal kidney lysates 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: 49 kDa
Observed band size: 50 kDa
Western blot - Anti-beta Tubulin antibody [EPR16774] - Loading Control (ab179513)
Beta Tubulin Western blot staining using rabbit Anti-beta Tubulin antibody
Blocking/Dilution buffer: 5% NFDM/TBST.
The amino acid sequence of ab179513 immunogen is identical to those of tubulin beta 2A, 2B, 3, 4A, 4B, 5, 6 and 8 at the same region.
All lanes: Western blot - Anti-beta Tubulin antibody [EPR16774] - Loading Control (ab179513) at 1/2000 dilution
Lane 1: Mouse brain lysates at 10 µg
Lane 2: Rat brain lysates at 10 µg
Lane 3: C6 (Rat glial tumor cells) whole cell lysates at 10 µg
Lane 4: RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates at 10 µg
Lane 5: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 10 µg
Lane 6: NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 49 kDa
Observed band size: 50 kDa
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody [EPR16774] (ab179513)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 cells labelling Mouse IgG1 with Mouse IgG1 monoclonal [R312-MouseIgG1]-Isotype control ab280974 at 1/20 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).
Confocal image showing no staining in RAW 264.7 cells.
Negative control 1: Mouse IgG1 monoclonal [R312-MouseIgG1]-Isotype control ab280974 at a 1/20 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 at a 1/1000 dilution.
Negative control 2: ab179513 at a 1/200 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 at a 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Tubulin antibody [EPR16774] (ab179513)
Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling beta Tubulin with ab179513 at 1/250 dilution, followed by Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasmic staining on neurons of human cerebral cortex is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
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 Tubulin antibody [EPR16774] (ab179513)
Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling beta Tubulin with ab179513 at 1/250 dilution, followed by Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasmic staining on tubules and the glomerulus of human kidney is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
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 Tubulin antibody [EPR16774] (ab179513)
Immunohistochemical analysis of paraffin-embedded Human glioma tissue labeling beta Tubulin with ab179513 at 1/250 dilution, followed by Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasmic staining on human glioma is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
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 Tubulin antibody [EPR16774] (ab179513)
Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling beta Tubulin with ab179513 at 1/250 dilution, followed by Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasm staining on neurons of mouse cerebral cortex is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
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 Tubulin antibody [EPR16774] (ab179513)
Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling beta Tubulin with ab179513 at 1/250 dilution, followed by Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasm staining on neurons of rat cerebral cortex is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody [EPR16774] (ab179513)
NIH/3T3 cells were fixed in 100% methanol and permeabilized with 0.1% Triton X-100. Primary antibody, Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade ab52484 at 1:2000 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 1/1000 dilution at RT for 45 min. ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade ab52484 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody [EPR16774] (ab179513)
RAW 264.7 cells were fixed in 4% PFA and permeabilized with 0.1% Triton X-100. Primary antibody, Anti-CD68 antibody [FA-11] ab53444 at 1:50 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-Rat secondary antibody (Goat Anti-Rat IgG H&L (Alexa Fluor® 488) ab150157) at 1/1000 dilution at RT for 45 min. ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-CD68 antibody [FA-11] ab53444 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI. Confocal image showing cytoplasmic staining in RAW 264.7 cell line.
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody [EPR16774] (ab179513)
Confocal image showing membranous staining in RAW 264.7 cells.

RAW 264.7, WEHI-231 and THP-1 cells were fixed in 4% PFA and permeabilized with 0.1% Triton X-100. Primary antibody, Anti-CD11b antibody [M1/70] ab8878 at 1:50 was incubated overnight at 4° C, followed by AlexaFluor® 488- conjugated Goat anti-Rat secondary antibody (Goat Anti-Rat IgG H&L (Alexa Fluor® 488) ab150157) at 1/1000 dilution at RT for 45 min. ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-CD11b antibody [M1/70] ab8878 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.

Negative control: WEHI-231 (PMID: 2457584)
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody [EPR16774] (ab179513)
Immunofluorescent analysis of 80% Methanol-fixed, 0.1% TritonX-100 permeabilized Jurkat (human T cell leukemia T lymphocyte) cells labeling SHP2 with Anti-SHP2 antibody [79/PTP1D/SHP2] ab290646 at 1/50 (20.18 µg/ml) dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (Green). Confocal image showing cytoplasmic staining in Jurkat cell line is observed. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 10 µg/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 µg/ml dilution.
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody [EPR16774] (ab179513)
MCF7/ HCT 116 cells were fixed in 4% PFA and permeabilized with 0.1% Triton X-100. Primary antibody, Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] ab70475 at 1:100 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-Mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113) at 1/1000 dilution at RT for 45 min. ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-Integrin beta 3 antibody [Y2/51] ab9509 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.

Confocal image showing membranous and cytoplasmic staining in MCF7 cells.
Negative control: HCT 116 (PMID: 14998492)
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody [EPR16774] (ab179513)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling Hsp27 with Anti-Hsp27 antibody [G3.1] ab2790 at 1/100 dilution, followed by AlexaFluor® 488-conjugated Goat anti-Mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113) at 1/1000 dilution at RT for 45 min. ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-Integrin beta 3 antibody [Y2/51] ab9509 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.