Anti-beta Tubulin antibody ab6046 is a rabbit polyclonal antibody that is used in beta Tubulin western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.
- Tried and trusted by researchers since 2004
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- Journal of cell science 136:2023Regulated dynamic subcellular GLUT4 localization revealed by proximal proteome mapping in human muscle cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAnuttoma Ray,Jennifer Wen,Lucie Yammine,Jeff Culver,Isabella Supardi Parida,Jeonifer Garren,Liang Xue,Katherine Hales,Qing Xiang,Morris J Birnbaum,Bei B Zhang,Mara Monetti,Timothy E McGrawPubMed 38126809
- PloS one 18:e02960852023ATXN2 is a target of N-terminal proteolysis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMonika Chitre,Patrick EmeryPubMed 38128014
- Nature communications 14:84202023Mutant GGGGCC RNA prevents YY1 from binding to Fuzzy promoter which stimulates Wnt/β-catenin pathway in C9ALS/FTD.Applications:Unspecified applicationReactive species:Unspecified reactive speciesZhefan Stephen Chen,Mingxi Ou,Stephanie Taylor,Ruxandra Dafinca,Shaohong Isaac Peng,Kevin Talbot,Ho Yin Edwin ChanPubMed 38110419
- Molecular medicine (Cambridge, Mass.) 29:1622023The linear ANRIL transcript P14AS regulates the NF-κB signaling to promote colon cancer progression.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWanru Ma,Junhua HuPubMed 38041015
- Cancer gene therapy 31:28-422023Downstream STING pathways IRF3 and NF-κB differentially regulate CCL22 in response to cytosolic dsDNA.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJihyun Kim,Jocelyn V Pena,Hannah P McQueen,Lingwei Kong,Dina Michael,Elmira M Lomashvili,Pamela R CookPubMed 37990062
- Scientific reports 13:204772023Mutant huntingtin confers cell-autonomous phenotypes on Huntington's disease iPSC-derived microglia.Applications:Unspecified applicationReactive species:Unspecified reactive speciesNina Stöberl,Jasmine Donaldson,Caroline S Binda,Branduff McAllister,Hazel Hall-Roberts,Lesley Jones,Thomas H Massey,Nicholas D AllenPubMed 37993517
- International journal of molecular sciences 24:2023Lipids Reduce Cytokine-Induced Pro-Inflammatory Signalling and Barrier Dysfunction in Human Keratinocyte Models.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJamie M L Kok,Georgina C Dowd,Jaydee D Cabral,Lyn M WisePubMed 38003573
- American journal of cancer research 13:5254-52702023PRDM16 from hepatic stellate cells-derived extracellular vesicles promotes hepatocellular carcinoma progression.Applications:Unspecified applicationReactive species:Unspecified reactive speciesChen Sun,Wenwen Xu,Yunhong Xia,Shuomin WangPubMed 38058806
- Reproductive biology and endocrinology : RB&E 21:1082023Thyroid hormone transport and metabolism are disturbed in the placental villi of miscarriage.Applications:Unspecified applicationReactive species:Unspecified reactive speciesZhen Yu,Xinghao Feng,Zhongshan Lin,Xuan Li,Shiyue Su,Huiru Cheng,Yuanyuan Yang,Zhaolian WeiPubMed 37968664
- Nature communications 14:73282023RNA mA methylation modulates airway inflammation in allergic asthma via PTX3-dependent macrophage homeostasis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesXiao Han,Lijuan Liu,Saihua Huang,Wenfeng Xiao,Yajing Gao,Weitao Zhou,Caiyan Zhang,Hongmei Zheng,Lan Yang,Xueru Xie,Qiuyan Liang,Zikun Tu,Hongmiao Yu,Jinrong Fu,Libo Wang,Xiaobo Zhang,Liling Qian,Yufeng ZhouPubMed 37957139
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| IHC-P | ICC/IF | IP | WB | |
|---|---|---|---|---|
| Human | Tested | Tested | Tested | Tested |
| Mouse | Expected | Expected | Expected | Tested |
| Rat | Expected | Expected | Expected | Tested |
| Chicken | Predicted | Predicted | Predicted | Predicted |
| Chinese hamster | Expected | Expected | Expected | Tested |
| Pig | Predicted | Predicted | Predicted | Predicted |
| Xenopus laevis | Predicted | Predicted | Predicted | Predicted |
| Zebrafish | Predicted | Predicted | Predicted | Predicted |
IHC-P
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 5 µg/mL | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Chinese hamster, Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Chicken, Pig, Xenopus laevis, Zebrafish | Dilution info - | Notes - |
ICC/IF
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 1 µg/mL | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Chinese hamster, Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Chicken, Pig, Xenopus laevis, Zebrafish | Dilution info - | Notes - |
IP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Chinese hamster, Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Chicken, Pig, Xenopus laevis, Zebrafish | Dilution info - | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Chinese hamster | Dilution info 1/500 | Notes - |
Species Mouse | Dilution info 1/500 | Notes - |
Species Rat | Dilution info 1/500 | Notes - |
Species Human | Dilution info 1/500 | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Chicken, Pig, Xenopus laevis, Zebrafish | Dilution info - | Notes - |
Associated Products
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Target data
Function
Tubulin is the major constituent of microtubules. It binds two moles of GTP, one at an exchangeable site on the beta chain and one at a non-exchangeable site on the alpha chain.
Alternative names
Tubulin beta chain, Tubulin beta-5 chain, OK/SW-cl.56, TUBB5, TUBB
Recommended products
Anti-beta Tubulin antibody ab6046 is a rabbit polyclonal antibody that is used in beta Tubulin western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.
- Tried and trusted by researchers since 2004
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.
- Purification technique
Affinity purification Immunogen
- Specificity
This antibody detects a single clean band at 50kD representing beta Tubulin. This band is significantly reduced by using peptide blocking.
- 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
Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.
If you do not find a host species to meet your needs, our catalogue and custom Chimeric range provides scientists the specificity of Abcam's RabMAbs in the species backbone of your choice. Remember to also review our range of edited cell lines, proteins and biochemicals relevant to your target that may help you further your research goals.
Abcam antibodies are extensively validated in a wide range of species and applications, so please check the reagent specifications meet your scientific needs before purchasing. If you have any questions or bespoke requirements, simply visit the Contact Us page to send us an inquiry or contact our Support Team ahead of purchase.
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
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.
38 product images
Immunoprecipitation - Anti-beta Tubulin antibody - Loading Control (ab6046)
Beta Tubulin was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Tubulin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab6046.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) ab99697).
Band: 50kDa: beta Tubulin.All lanes: Immunoprecipitation - Anti-beta Tubulin antibody - Loading Control (ab6046)
Predicted band size: 49 kDa
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
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 ab6046 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) preadsorbed ab186697 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046) 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® 790) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) preadsorbed ab186697) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 50 kDa
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
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 ab6046 overnight at 4°C. Antibody binding was detected using Goat Anti-Guinea pig IgG H&L (Alexa Fluor® 750) ab175758 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046) at 1 µg/mL
All lanes: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
SecondaryAll lanes: Western blot - Goat Anti-Guinea pig IgG H&L (Alexa Fluor® 750) (Goat Anti-Guinea pig IgG H&L (Alexa Fluor® 750) ab175758) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 50 kDa
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
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 ab6046 overnight at 4°C. Antibody binding was detected using Donkey F(ab')2 Anti-Rabbit IgG H&L (Alexa Fluor® 750) preadsorbed ab175730 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046) at 1 µg/mL
All lanes: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
SecondaryAll lanes: Western blot - Donkey F(ab')2 Anti-Rabbit IgG H&L (Alexa Fluor® 750) preadsorbed (Donkey F(ab')2 Anti-Rabbit IgG H&L (Alexa Fluor® 750) preadsorbed ab175730) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 50 kDa
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
All lanes: Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046) at 1/500 dilution
Lanes 1 and 5: HeLa Cell lysate at 20 µg
Lanes 2 and 6: A431 Cell lysate at 20 µg
Lanes 3 and 7: MCF7 Cell lysate at 20 µg
Lanes 4 and 8: 293 Cell lysate at 20 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 49 kDa
Observed band size: 55 kDa
Exposure time: 10s
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
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 ab6046 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG Fc (Alexa Fluor® 750) ab175735 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046) 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 Fc (Alexa Fluor® 750) (Goat Anti-Rabbit IgG Fc (Alexa Fluor® 750) ab175735) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 50 kDa
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
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 ab6046 overnight at 4°C. Antibody binding was detected using Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 750) ab175731 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046) 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® 750) (Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 750) ab175731) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 50 kDa
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
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 ab6046 overnight at 4°C. Antibody binding was detected using Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 750) preadsorbed ab175728 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046) 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® 750) preadsorbed (Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 750) preadsorbed ab175728) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 37 kDa, 50 kDa
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
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 ab6046 overnight at 4°C. Antibody binding was detected using Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 790) preadsorbed ab186693 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046) at 1 µg/mL
All lanes: HeLa Whole Cell Lysate at 10 µg
SecondaryAll lanes: Western blot - Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 790) preadsorbed (Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 790) preadsorbed ab186693) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 50 kDa
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
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 ab6046 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 750) ab175732 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046) 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® 750) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 750) ab175732) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 37 kDa, 50 kDa
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
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 ab6046 overnight at 4°C. Antibody binding was detected using Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 680) preadsorbed ab186692 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046) 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® 680) preadsorbed (Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 680) preadsorbed ab186692) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 50 kDa
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
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 ab6046 overnight at 4°C. Antibody binding was detected using Donkey Anti-Sheep IgG H&L (Alexa Fluor® 750) ab175756 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046) at 1 µg/mL
All lanes: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
SecondaryAll lanes: Donkey anti-Sheep IgG H&L (AlexaFluor750) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 94 kDa
Observed band size: 15 kDa, 50 kDa, 94 kDa
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
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 ab6046 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 750) preadsorbed ab175733 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046) 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® 750) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 750) preadsorbed ab175733) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 50 kDa
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
ab6046 staining beta 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 ab6046 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.
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
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 ab6046 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 680) preadsorbed ab186696 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
All lanes: Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046) 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) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 680) preadsorbed ab186696) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 50 kDa
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
All lanes: Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046) at 1 µg/mL
Lane 1: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg
Lane 2: NIH3T3 (Mouse embryo fibroblast cell line) whole cell lysate at 10 µg
Lane 3: PC12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg
Lane 4: CHO/K1 (Chinese hamster ovary cell line) whole cell lysate at 10 µg
SecondaryAll lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 49 kDa
Observed band size: 51 kDa
Exposure time: 30s
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
ICC/IF image of ab6046 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min), 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 1h to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6046, 1μg/ml) overnight at +4°C. The secondary antibody (green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. 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 100% methanol (5 min).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Tubulin antibody - Loading Control (ab6046)
IHC image of beta Tubulin staining in human liver carcinoma FFPE section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab6046, 5µg/ml, for 15 mins 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.
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
alpha smooth muscle Actin immunofluorescence staining of HeLa cells using mouse anti-alpha smooth muscle Actin antibody
Anti-alpha smooth muscle Actin antibody [1A4] ab7817 staining alpha smooth muscle Actin in wild-type HeLa cells (top panel) and ACTA2 knockout HeLa cells (Human ACTA2 knockout HeLa cell line ab264014) (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% 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 with Anti-alpha smooth muscle Actin antibody [1A4] ab7817 at 5ug/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2 ug/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 2 ug/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
Ctip2 immunofluorescence staining of Jurkat and Daudi cells using rat anti-Ctip2 antibody
Immunofluorescence staining of Ctip2 using Anti-Ctip2 antibody [25B6] ab18465 in Jurkat cells (+ve expression control top panel) and Daudi cells (-ve expression control bottom panel). The cells were fixed with 4% formaldehyde (10 min) permeabilized with 0.1% 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 with Anti-Ctip2 antibody [25B6] ab18465 at 0.2 µg/mL and ab6046 at 1 µg/mL overnight at +4°C followed by a further incubation at room temperature for 1h with Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed ab150165) (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150084) (shown in red) both at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
Ctip2 immunofluorescence staining of Jurkat and Daudi cells using rat anti-Ctip2 antibody
Immunofluorescence staining of Ctip2 using Anti-Ctip2 antibody [25B6] ab18465 in Jurkat cells (+ve expression control top panel) and Daudi cells (-ve expression control bottom panel). The cells were fixed with 100% methanol (5 min) permeabilized with 0.1% 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 with Anti-Ctip2 antibody [25B6] ab18465 at 0.2 µg/mL and ab6046 at 1 µg/mL overnight at +4°C followed by a further incubation at room temperature for 1h with Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed ab150165) (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150084) (shown in red) both at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
Anti-PAX6 antibody [AD2.38] ab78545 staining Pax6 in primary mouse neurons/glia, DIV1 (top row) and DIV14 (bottom row) both prepared from E18 mouse hippocampal brain area (obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. C57EHP). 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 Anti-PAX6 antibody [AD2.38] ab78545 at 5 µg/ml and ab6046, rabbit polyclonal to beta Tubulin, at 1/1000 dilution. Cells were then incubated with with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150084, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
A subset of cells showed staining in the nucleus at DIV1 (likely neuroprogenitor cells), while differentiated neurons (DIV14) where Pax6 negative.
The antibody was not suitable to detect Pax6 in cells fixed with 100% methanol (5 min).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
Immunofluorescence staining of E-Cadherin using Anti-E Cadherin antibody [4A2] ab231303 in wild-type A431 cells (top panel) and CDH1 knockout A431 cells (bottom panel). The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% 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 with Anti-E Cadherin antibody [4A2] ab231303 at 1.0 µg/mL and ab6046 at 1 µg/mL overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150084) (shown in red), both at 1/1000. Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a confocal section is shown.
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
Immunofluorescence staining of E-Cadherin using Anti-E Cadherin antibody [4A2] ab231303 in wild-type A431 cells (top panel) and CDH1 knockout A431 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% 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 with Anti-E Cadherin antibody [4A2] ab231303 at 1.0 µg/mL and ab6046 at 1 µg/mL overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150084) (shown in red), both at 1/1000. Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a confocal section is shown.
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
Anti-DDX4 / MVH antibody [mAbcam27591] ab27591 staining DDX4/MVH in mES cells. The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% 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 with Anti-DDX4 / MVH antibody [mAbcam27591] ab27591 at 10μg/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
Anti-VPS35 antibody [2D3] ab57632 staining VPS35 in wild-type HAP1 cells (top panel) and VPS35 knockout HAP1 cells (bottom panel). The cells were fixed with 4% PFA (10 min), permeabilized with 0.1% 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 with Anti-VPS35 antibody [2D3] ab57632 at 0.2 µg/mL and ab6046 at 1 µg/mL overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Mouse IgG (Alexa Fluor® 488) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) (shown in pseudo colour green) and goat secondary antibody to Rabbit IgG (Alexa Fluor® 594) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150084) (shown in pseudo colour red) both at 1/1000. Nuclear DNA was labelled with DAPI (shown in pseudo colour blue).
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-beta Tubulin antibody - Loading Control (ab6046)
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
Cells incubated overnight at 4°C. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h.
Nuclear DNA was labelled with DAPI (shown in blue). 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-beta Tubulin antibody - Loading Control (ab6046)
Anti-p53 antibody [DO-1] - ChIP Grade ab1101 staining wild-type p53 in MCF7 cells (a low expressing cell line). The cells were fixed with 100% methanol (5 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 Anti-p53 antibody [DO-1] - ChIP Grade ab1101 at 0.2µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 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-beta Tubulin antibody - Loading Control (ab6046)
Anti-p53 antibody [DO-1] - ChIP Grade ab1101 staining wild-type p53 in Hek293 cells (a high expressing cell line). The cells were fixed with 100% methanol (5 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 Anti-p53 antibody [DO-1] - ChIP Grade ab1101 at 0.2µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 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-beta Tubulin antibody - Loading Control (ab6046)
Anti-p53 antibody [DO-1] - ChIP Grade ab1101 staining mutant p53 in A431 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 Anti-p53 antibody [DO-1] - ChIP Grade ab1101 at 0.2µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) 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-beta Tubulin antibody - Loading Control (ab6046)
Anti-F4/80 antibody [F4/80] - Macrophage Marker ab90247 staining F4/80 in Raw264.7 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween 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 Anti-F4/80 antibody [F4/80] - Macrophage Marker ab90247 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed ab150165, Goat polyclonal Secondary Antibody to Rat IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
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-beta Tubulin antibody - Loading Control (ab6046)
Anti-HIF-1 alpha antibody [ESEE122] ab8366 staining HIF-1 alpha in Hela cells. Untreated and BrdU treated (10µM for 24 hours) 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 Anti-HIF-1 alpha antibody [ESEE122] ab8366 at 10µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
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