Anti-alpha Tubulin antibody - Microtubule Marker (ab18251) is a rabbit polyclonal antibody detecting alpha Tubulin in Western Blot, Flow Cytometry (Intra), Flow Cytometry, ICC/IF. Suitable for Human, Mouse, Rat.
- Over 360 publications
- Trusted since 2005
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- EMBO reports 24:e574402023Metabolic plasticity sustains the robustness of Caenorhabditis elegans embryogenesis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSiyu Chen,Xing Su,Jinglin Zhu,Long Xiao,Yulin Cong,Leilei Yang,Zhuo Du,Xun HuangPubMed 37885348
- Nature communications 14:49712023High-sensitive nascent transcript sequencing reveals BRD4-specific control of widespread enhancer and target gene transcription.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAnnkatrin Bressin,Olga Jasnovidova,Mirjam Arnold,Elisabeth Altendorfer,Filip Trajkovski,Thomas A Kratz,Joanna E Handzlik,Denes Hnisz,Andreas MayerPubMed 37591883
- Frontiers in pharmacology 14:11698122023Stimulatory effects of on human melanocyte proliferation, migration, and melanogenesis: and studies.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKhalid Alghamdi,Zeyad Alehaideb,Ashok Kumar,Hamad Al-Eidi,Sahar S Alghamdi,Rasha Suliman,Rizwan Ali,Feras Almourfi,Saleh M Alghamdi,Mohamed Boudjelal,Sabine Matou-NasriPubMed 37197407
- Molecular biology of the cell 34:ar632023Cortical dynein drives centrosome clustering in cells with centrosome amplification.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDayna L Mercadante,William A Aaron,Sarah D Olson,Amity L ManningPubMed 37017483
- The EMBO journal 42:e1135192023Functionally distinct promoter classes initiate transcription via different mechanisms reflected in focused versus dispersed initiation patterns.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLeonid Serebreni,Lisa-Marie Pleyer,Vanja Haberle,Oliver Hendy,Anna Vlasova,Vincent Loubiere,Filip Nemčko,Katharina Bergauer,Elisabeth Roitinger,Karl Mechtler,Alexander StarkPubMed 37013908
- Scientific reports 13:53662023Heat denaturation enables multicolor X10-STED microscopy.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKim Ann Saal,Ali H Shaib,Nikolaos Mougios,Dagmar Crzan,Felipe Opazo,Silvio O RizzoliPubMed 37005431
- ACS bio & med chem Au 3:261-2692023Live-Cell SOFI Correlation with SMLM and AFM Imaging.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRiley B Hargreaves,Sam Duwé,Ashley M Rozario,Alison M Funston,Rico F Tabor,Peter Dedecker,Donna R Whelan,Toby D M BellPubMed 37363082
- Nature communications 14:17152023Distinct dynein complexes defined by DYNLRB1 and DYNLRB2 regulate mitotic and male meiotic spindle bipolarity.Applications:Unspecified applicationReactive species:Unspecified reactive speciesShuwen He,John P Gillies,Juliana L Zang,Carmen M Córdoba-Beldad,Io Yamamoto,Yasuhiro Fujiwara,Julie Grantham,Morgan E DeSantis,Hiroki ShibuyaPubMed 36973253
- Journal of virology 97:e01758222023YTHDF2 Is Downregulated in Response to Host Shutoff Induced by DNA Virus Infection and Regulates Interferon-Stimulated Gene Expression.Applications:Unspecified applicationReactive species:Unspecified reactive speciesCharles R Hesser,Derek WalshPubMed 36916936
- International journal of molecular sciences 24:2023Selectivity of Hydroxamate- and Difluoromethyloxadiazole-Based Inhibitors of Histone Deacetylase 6 In Vitro and in Cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJakub Ptacek,Ivan Snajdr,Jiri Schimer,Zsofia Kutil,Jana Mikesova,Petra Baranova,Barbora Havlinova,Werner Tueckmantel,Pavel Majer,Alan Kozikowski,Cyril BarinkaPubMed 36902164
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA- Form
- Liquid
- Clonality
- Polyclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Consider this alternative
Reactivity data
Select an application| Flow Cyt (Intra) | WB | ICC/IF | |
|---|---|---|---|
| Human | Tested | Tested | Tested |
| Mouse | Tested | Tested | Tested |
| Rat | Tested | Tested | Tested |
| African green monkey | Predicted | Predicted | Predicted |
| Chicken | Predicted | Predicted | Predicted |
| Chinese hamster | Predicted | Predicted | Predicted |
| Cow | Predicted | Predicted | Predicted |
| Drosophila melanogaster | Predicted | Predicted | Predicted |
| Indian muntjac | Predicted | Predicted | Predicted |
Flow Cyt (Intra)
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info 0.01 µg for 105 Cells | Notes - |
Species Rat | Dilution info 0.01 µg for 105 Cells | Notes - |
Species Human | Dilution info 0.01 µg for 105 Cells | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Chicken, Cow, Drosophila melanogaster, Indian muntjac, African green monkey, Chinese hamster | Dilution info - | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info 0.5 µg/mL | Notes Abcam recommends using milk as the blocking agent. |
Species Rat | Dilution info 0.5 µg/mL | Notes Abcam recommends using milk as the blocking agent. |
Species Human | Dilution info 0.5 µg/mL | Notes Abcam recommends using milk as the blocking agent. |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Chicken, Cow, Drosophila melanogaster, Indian muntjac, African green monkey, Chinese hamster | Dilution info - | Notes - |
ICC/IF
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info 1-5 µg/mL | Notes - |
Species Rat | Dilution info 1-5 µg/mL | Notes - |
Species Human | Dilution info 1-5 µg/mL | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Chicken, Cow, Drosophila melanogaster, Indian muntjac, African green monkey, Chinese hamster | Dilution info - | Notes - |
Associated Products
Select an associated product type
6 products for Alternative Product
Target data
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
TUBA1, TUBA4A, Tubulin alpha-4A chain, Alpha-tubulin 1, Testis-specific alpha-tubulin, Tubulin H2-alpha, Tubulin alpha-1 chain
Recommended products
Anti-alpha Tubulin antibody - Microtubule Marker (ab18251) is a rabbit polyclonal antibody detecting alpha Tubulin in Western Blot, Flow Cytometry (Intra), Flow Cytometry, ICC/IF. Suitable for Human, Mouse, Rat.
- Over 360 publications
- Trusted since 2005
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA- Form
- Liquid
- Clonality
- Polyclonal
- Immunogens
- The exact immunogen used to generate this antibody is proprietary information.
- Purification technique
- Affinity purification Immunogen
- Specificity
Replenishment batches of our polyclonal antibody, ab18251 are tested in WB. Previous batches were additionally validated in Flow Cyt and ICC/IF. These applications are still expected to work and are covered by our Abpromise guarantee. You may also be interested in our alternative recombinant antibody, ab52866.
- 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
Anti-alpha Tubulin antibody - Microtubule Marker (ab18251) is a rabbit polyclonal antibody and is validated for use in Flow Cyt (Intra), ICC/IF, WB in human, mouse, rat samples.
alpha Tubulin antibodies are often used as loading controls in Western Blot. Anti-alpha Tubulin antibody - Microtubule Marker has been verified in Western Blot samples and detects a band at 50kDa Molecular weight.
Anti-alpha Tubulin antibody - Microtubule Marker (ab18251) specifically detects alpha Tubulin (UniProt ID: P68366; Molecular weight: 50kDa) and is sold in 50 µg selling sizes.
Quality and Validation
Abcam's high quality validation processes ensure Anti-alpha Tubulin antibody - Microtubule Marker (ab18251) has high sensitivity and specificity.
Anti-alpha Tubulin antibody - Microtubule Marker (ab18251) has been cited over 362 times in peer reviewed journals and is trusted by the scientific community.
Anti-alpha Tubulin antibody - Microtubule Marker (ab18251) has 21 independent reviews from customers.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our 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.
15 product images
Western blot - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
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 ab18251 overnight at 4°C. Antibody binding was detected using Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 680) ab175772 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-alpha Tubulin antibody - Loading Control (ab18251) 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 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
SecondaryAll lanes: Western blot - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 680) (Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 680) ab175772) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 52 kDa
Western blot - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Western blot staining of HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) using rabbit Anti-alpha Tubulin antibody
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 ab18251 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) ab175781 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-alpha Tubulin antibody - Loading Control (ab18251) at 1 µg/mL
All lanes: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 10 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) ab175781) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 50 kDa
Western blot - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Western blot staining of HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate using rabbit Anti-alpha Tubulin antibody
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 ab18251 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 680) ab175773 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-alpha Tubulin antibody - Loading Control (ab18251) at 1 µg/mL
All lanes: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 680) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 680) ab175773) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 50 kDa
Western blot - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Western blot staining of HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate using rabbit Anti-alpha Tubulin antibody
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 ab18251 overnight at 4°C. Antibody binding was detected using Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 790) ab175780 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-alpha Tubulin antibody - Loading Control (ab18251) at 1 µg/mL
All lanes: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
SecondaryAll lanes: Western blot - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 790) (Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 790) ab175780) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 50 kDa
Flow Cytometry (Intracellular) - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Flow Cytometry (Intracellular) staining using rabbit Anti-alpha Tubulin antibody
Overlay histogram showing NIH3T3 cells stained with ab18251 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 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 (ab18251, 0.01μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (polyclonal) (ab27478, 0.01μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Flow Cytometry (Intracellular) - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Flow Cytometry (Intracellular) staining using rabbit Anti-alpha Tubulin antibody
Overlay histogram showing SV40LT-SMC cells stained with ab18251 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 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 (ab18251, 0.01μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (polyclonal) (ab27478, 0.01μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Flow Cytometry (Intracellular) - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Flow Cytometry (Intracellular) staining using rabbit Anti-alpha Tubulin antibody
Overlay histogram showing Caco2 cells stained with ab18251 (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 (ab18251, 0.01μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (polyclonal) (ab27478, 0.01μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Western blot - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
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 Licor blocking buffer before being incubated with ab18251 overnight at 4°C. Antibody binding was detected using Anti-Rabbit Alexa Fluor® 790 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) ab175781) at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-alpha Tubulin antibody - Loading Control (ab18251) at 0.5 µg/mL
Lane 1: HeLa (Human epithelial carcinoma cell line) whole cell lysate at 20 µg
Lane 2: HEK-293 (Human embryonic kidney cell line) whole cell lysate at 20 µg
Lane 3: HepG2 (Human hepatocellular liver carcinoma cell line) whole cell lysate at 20 µg
Lane 4: Caco-2 (Human colonic carcinoma cell line) whole cell lysate at 20 µg
Lane 5: NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 20 µg
Lane 6: PC-12 (Rat adrenal pheochromocytoma cell line) whole cell lysate at 20 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) ab175781) at 1/10000 dilution
Predicted band size: 27 kDa, 50 kDa
Observed band size: 44 kDa, 52 kDa
Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-alpha Tubulin antibody
ab18251 staining alpha-Tubulin in SV40LT-SMC cells.
The cells were fixed with 100% methanol for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated with ab18251 at 1 μl/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1 μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1 hour with an anti-rabbit Alexa Fluor® 488 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and anti-mouse Alexa Fluor® 594 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labeled in blue with DAPI.
Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-alpha Tubulin antibody
ab18251 staining alpha-Tubulin in NIH3T3 cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab18251 at 1 μl/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-rabbit Alexa Fluor® 488 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and anti-mouse Alexa Fluor® 594 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-alpha Tubulin antibody
ab18251 staining alpha-Tubulin in Caco-2 cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab18251 at 5 μl/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1 μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-rabbit Alexa Fluor® 488 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and anti-mouse Alexa Fluor® 594 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-alpha Tubulin antibody
ab18251 staining alpha 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 ab18251 at 1µg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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).
Also suitable in cells fixed with 100% methanol (5 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Immunocytochemistry/ Immunofluorescence - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-alpha Tubulin antibody
ab18251 at a 1/8000 dilution staining human HeLa cells by immunocytochemistry. The cells were paraformaldehyde fixed and incubated with the antibody for 30 minutes. The secondary antibody was a Cy3® conjugated Goat Anti-Rabbit IgG (H+L). The image shows staining of an interphase IM cell.
This image is courtesy of an Abreview by Kirk McManus submitted on 27 February 2006.
Western blot - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
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 ab18251 overnight at 4°C. Antibody binding was detected using an anti-rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051), and visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406
All lanes: Western blot - Anti-alpha Tubulin antibody - Loading Control (ab18251) at 1 µg/mL
Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Lane 2: A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 50 kDa
Exposure time: 1min
Western blot - Anti-alpha Tubulin antibody - Loading Control (ab18251)
Alpha Tubulin Western blot staining using rabbit Anti-alpha Tubulin antibody
ab18251 detects a strong band at 50 kDa corresponding to alpha tubulin. Cross-reactivity is also seen with other lower molecular weight bands. This may be reduced by using the antibody at a lower working concentration.
All lanes: Western blot - Anti-alpha Tubulin antibody - Loading Control (ab18251) at 0.5 µg/mL
Lane 1: HeLa lysate at 20 µg
Lane 2: A431 lysate at 20 µg
SecondaryAll lanes: Alexa Flour Goat polyclonal to Rabbit IgG (700) at 1/10000 dilution
Predicted band size: 50 kDa
Observed band size: 30 kDa, 50 kDa
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