alpha Tubulin [ac Lys40] Antibody (6-11B-1)

Immunohistochemistry: alpha Tubulin [ac Lys40] Antibody (6-11B-1) [NB600-567] - Nerve pathways in the embryonic zebrafish brain and spinal cord revealed by alpha Tubulin (ac Lys40) Antibody (6-11B-1). Immunoperoxidase staining of a whole-mount embyro at 24 hours of development.

Western Blot: alpha Tubulin [ac Lys40] Antibody (6-11B-1) [NB600-567] - Western Blot validation of monoclonal anti-Acetylated Tubulin antibody clone: 6-11B-1 at 1:2000 dilution with goat anti-Mouse IgG-Peroxidase secondary antibodychemiluminescent substrate development. The lysates employed were from cell lines: 1. HeLa, 2. U87, 3. COS7, 4. P19, 5. RAT2, 6. CHO, 7. MDBK and 8. MDCK Alpha tubulin molecular weight: 50 kDa.

Immunocytochemistry/Immunofluorescence: alpha Tubulin [ac Lys40] Antibody (6-11B-1) [NB600-567] - HeLa cells were fixed with 4% paraformaldehyde followed permeabilization with 0.5% Triton X-100. Fixed cells were stained with 1:2,000 monoclonal Tubulin, Acetylated antibody Clone: 6-11B-1 . The antibody was detected using goat anti-Mouse IgG, Cy3 conjugate followed by counterstaining with DAPI (blue).

Immunoprecipitation: alpha Tubulin [ac Lys40] Antibody (6-11B-1) [NB600-567] - Identification of a surrogate peptide YCLEHGIQPDGQMPSDK40(ac)TIGGGDDSF with mass spectrometry. This peptide was immunoprecipitated with 6-11B-1 mAb from peptic digested rat brain tissue lysate. Image submitted via customer review.

Product Discontinued

alpha Tubulin [ac Lys40] Antibody (6-11B-1) Summary

• Immunogen: This alpha Tubulin [ac Lys40] Antibody (6-11B-1) was developed against acetylated tubulin from the outer arm of Strongylocentrotus purpuratus (sea urchin)
• Modification: ac Lys40
• Localization: Centrosomal.
• Marker: Microtubule Marker
• Specificity: This alpha Tubulin [ac Lys40] Antibody (6-11B-1) recognizes an epitope located on the alpha 3 isoform of Chlamydomonas axonemal alpha-tubulin, within four residues of Lys-40 when this amino acid is acetylated
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: TUBA1A
• Purity: Unpurified

Applications/Dilutions

• Dilutions:
  • Dot Blot 1:100-1:2000
  • Electron Microscopy 1:10-1:500
  • ELISA 1:100-1:2000
  • Immunocytochemistry/ Immunofluorescence 1:10-1:2000
  • Immunohistochemistry 1:10-1:500
  • Immunoprecipitation
  • Radioimmunoassay 1:100-1:2000
  • Western Blot 1:2000
• Theoretical MW: 50 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Ascites
• Preservative: 0.09% Sodium Azide
• Purity: Unpurified

Alternate Names for alpha Tubulin [ac Lys40] Antibody (6-11B-1)

• alpha tubulin loading control
• alpha tubulin monoclonal
• alpha Tubulin
• Alpha-Tubulin 3
• B-ALPHA-1
• FLJ25113
• hum-a-tub1
• hum-a-tub2
• LIS3
• TUBA1 monoclonal
• TUBA1A
• TUBA3
• tublin
• tubulin alpha-1A chain
• Tubulin alpha-3 chain
• Tubulin B-Alpha-1
• tubulin monoclonal
• tubulin, alpha 1a
• tubulin, alpha 3
• tubulin, alpha, brain-specific

Background

Tubulin is the major building block of microtubules. Microtubules function as structural and mobile elements in mitosis, intracellular transport, flagellar movement and in the cytoskeleton. The tubulin superfamily is composed of six distinct families. Genes from the alpha, beta and gamma tubulin families are found in all eukaryotes. Microtubules of the eukaryotic cytoskeleton perform essential and diverse functions and are composed of a heterodimer of alpha and beta tubulins. Alpha and beta tubulin dimers are assembled to 13 protofilaments that form a microtubule of 22 nm diameter with a theoretical molecular weight of 50 kDa.

Tyrosine ligase adds a C-terminal tyrosine to monomeric alpha tubulin. Assembled microtubules can again be detyrosinated by a cytoskeleton associated carboxypeptidase. Detyrosinated alpha tubulin is referred to as Glu-tubulin. Another post-translational modification of detyrosinated alpha tubulin is C-terminal polyglutamylation which is characteristic for microtubules in neuronal cells and the mitotic spindle.

Like GAPDH and beta-actin, alpha/beta tubulin is often used as a loading control in immunoblot applications (1). Alpha/beta tubulin is also good for counterstaining microtubules in immunofluorescence (2).

References

1. Hannen, R., Selmansberger, M., Hauswald, M., Pagenstecher, A., Nist, A., Stiewe, T., . . . Bartsch, J. W. (2019). Comparative Transcriptomic Analysis of Temozolomide Resistant Primary GBM Stem-Like Cells and Recurrent GBM Identifies Up-Regulation of the Carbonic Anhydrase CA2 Gene as Resistance Factor. Cancers (Basel), 11(7). doi:10.3390/cancers11070921

2. Nel, M., Joubert, A. M., Dohle, W., Potter, B. V., & Theron, A. E. (2018). Modes of cell death induced by tetrahydroisoquinoline-based analogs in MDA-MB-231 breast and A549 lung cancer cell lines. Drug Des Devel Ther, 12, 1881-1904. doi:10.2147/dddt.S152718

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for alpha Tubulin Antibody (NB600-567)(2)

Publications using NB600-567

• Yang X, Naughton S, Ha Z et al. Mass Spectrometric Quantitation of Tubulin Acetylation from Pepsin-Digested Rat Brain Tissue Using a Novel Stable Isotope Standard and Capture by Anti-Peptide Antibody (SISCAPA) Method. Anal Chem 2018-01-24 [PMID: 29320166] (IP, Rat)
• Mardilovich K, Gabrielsen M et al. Elevated LIM kinase 1 in nonmetastatic prostate cancer reflects its role in facilitating androgen receptor nuclear translocation. Mol Cancer Ther 2015-01-01 [PMID: 25344584] (ICC/IF, Human)

Review for alpha Tubulin Antibody (NB600-567) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Rat.

reviewed by: Xiangkun Yang IP Rat 01/20/2018

Summary

• Application: Immunoprecipitation
• Sample Tested: Rat brain lysate (ventral hippocampus)
• Species: Rat
• Lot: didn't keep the original container

Comments

• Comments: This is an application not reported before. I have sent my publication including this application a couple of days ago.

FAQs for alpha Tubulin Antibody (NB600-567). (Showing 1 - 4 of 4 FAQs).

1. The data sheet for Tubulin [ac Lys40] antibody (6-11B-1) (catalog number for Novus Biologicals: NB600-567) indicates that this antibody works in ELISA and I would like to request either the protocol or the literature reference for it at your earliest convenience. Do you also have the information on the capture antibody that you used and the input quantity available?
   • Here is a link to our ELISA protocol. NB600-567 was tested and validated for use in an indirect ELISA, and as such was used as the detection antibody. For the testing of this antibody, the ELISA plate wells were coated with the immunogen, acetylated a-tubulin from the outer arm of sea urchin sperm axonemes, and no capture antibody was used. Unfortunately I don't have any information regarding the input quantity, but please let me know if you have additional questions.
2. We are looking for an antibody which is a specific marker for Mouse Bronchial cilia in IHC(p) technique. Preferably not from a mouse host. Can you help?
   • According to the literature I have reviewed on bronchial cilia markers, the most commonly used marker is acetylated alpha tubulin (AcTub). Please see the relevant publications with images attached (Song et al. 2014, Schamberger et al. 2014). We have one antibody to AcTub that is validated for IHC-P and mouse (it is however also raised in mouse, hence mouse on mouse blocking step would be necessary): NB600-567. There are also some reports of using beta tubulin as cilial marker (Davis et al. 2015). For rabbit host antibodies, you may look at our best seller products like NB600-936 and NB100-56459
3. Will this alpha tubulin antibody recognize both isoforms of alpha tubulin? 
   • The epitope for this alpha tubulin antibody lies on the C-terminus of the protein and the difference between the two major isoforms is within the first 35 aa of the N-terminus so this alpha tubulin antibody will recognize both isoforms.
4. We would like to stain cilia with an acetylated alpha tubulin antibody in our cells, but I am unsure if this antibody will be able to conclusively differentiate cilia from other structures such as spindle pole bodies. Does anyone know what acetylated alpha tubulin antibodies might bind to apart from cilia?
   • Acetylated alpha tubulin is found in relatively stable microtubules. It is best practice to use this marker together with a centrosome/centriole marker, which will stain the basal bodies at the base of the cilium. After that, it is relatively straightforward to identify the acetylated alpha tubulin signal that corresponds to the cilium.

Bioinformatics

• Gene Symbol: TUBA1A
• Entrez:
  • 7846(Human)
  • 22142(Mouse)
  • 64158(Rat)