alpha Tubulin 4a Antibody (B-5-1-2) - BSA Free

Western Blot: alpha Tubulin 4a Antibody (B-5-1-2) - BSA Free [NB120-11304] - Cell line lysates were separated on SDS-PAGE and probed with Monoclonal Anti-alpha-Tubulin Clone: B-5-1-2. The antibody was developed using Goat Anti-Mouse IgG-Peroxidase and a chemiluminescent substrate. Lanes: 1.) HeLa, 2.) JURKAT, 3.) COS7, 4.) NIH-3T3, 5.) PC-12, 6.) RAT2, 7.) CHO, 8.) MDBK, 9.) MDCK

Immunocytochemistry/Immunofluorescence: alpha Tubulin 4a Antibody (B-5-1-2) [NB120-11304] - Staining of HEK293T cells transfected with a recombinant protein HA tagged, detected with Anti-HA Mouse primary antibody and Anti-Mouse Alexa543 secondary antibody. Microtubules detected with Anti-alpha-tubulin Mouse primary antibody and Anti-Mouse Alexa488 secondary antibody.

Immunocytochemistry/Immunofluorescence: alpha Tubulin 4a Antibody (B-5-1-2) [NB120-11304] - CFB cells were fixed and permeabilized with methanol followed by acetone. Fixed cells were stained with Monoclonal Anti-alpha-Tubulin antibody produced in mouse clone No. B-5-1-2. The antibody was developed using Anti-Mouse IgG (Fab specific)-FITC antibody produced in goat. Cells were counterstained with DAPI (blue) to stain nuclei.

alpha Tubulin 4a Antibody (B-5-1-2) - BSA Free [NB120-11304] - Chicken fibroblasts cells were fixed and permeabilized with methanol followed by acetone. Fixed cells were stained with 1 ug/mL Anti-alpha-Tubulin antibody, Mouse monoclonal, Clone: DM1A (A). The antibody was developed using 1:40 Goat Anti-Mouse IgG (Fab specific)-FITC antibody and 1 ug/mL Monoclonal Anti-alpha-Tubulin antibody produced in Mouse, Clone: B-5-1-2 (B). The antibody was developed using 1:40 Goat Anti-Mouse IgG (Fab specific)-FITC antibodyResults:Two Anti-alpha-Tubulin antibodies, T6199 (A) and T6074 (B) target different regions of alpha-Tubulin show similar staining profiles between the two antibodies, demonstrating Independent Antibody Verification.

• Reactivity: Hu, Mu, Rt, Bv, Ch, Pm
• Applications: WB, Simple Western, ICC/IF, IP, MiAr
• Clone: B-5-1-2
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 2.0 mg/ml

alpha Tubulin 4a Antibody (B-5-1-2) - BSA Free Summary

• Immunogen: Sarkosyl-resistant filaments from Stronglycentrotus purpuratus (sea urchin).
• Localization: Cytoplasmic
• Specificity: Recognizes an epitope located at the C-terminal end of the alpha-tubulin isoform in a variety of organisms.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: TUBA4A
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/Immunofluorescence 0.5-1 ug/ml
  • Immunoprecipitation 1:10-1:500
  • Microarray
  • Simple Western
  • Western Blot 0.25-0.5 ug/ml
• Application Notes: ICC: Concentration is recommended using cultured chicken fibroblasts (CFB). WB: Concentration is recommended using a total cell extract of the human foreskin fibroblast cell line (FS11). Predicted molecular weight: 50 kDa.
  Simple Western reported by an internal validation. Separated by Size-All, antibody dilution of 1:50; matrix was 12-230 kDa.
• 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.
• Control:

  Tubulin alpha-1B Overexpression Lysate

Reactivity Notes

Cross-reacts with African Green Monkey, Chlamydomonas reinhardtii, kangaroo rat and Sea Urchin. Not yet tested in other species. 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: 10mM PBS (pH 7.4)
• Preservative: 15mM Sodium Azide
• Concentration: 2.0 mg/ml
• Purity: Protein A or G purified

Alternate Names for alpha Tubulin 4a Antibody (B-5-1-2) - BSA Free

• alpha 1 (testis specific)
• alpha Tubulin 4a
• Alpha-tubulin 1
• FLJ30169
• H2-ALPHA
• Testis-specific alpha-tubulin
• TUBA1
• TUBA4A
• Tubulin alpha-1 chain
• tubulin alpha-4A chain
• Tubulin H2-alpha
• tubulin, alpha 1
• tubulin, alpha 4a

Background

Microtubules are involved in a wide variety of cellular activities ranging from mitosis and transport events to cell movement and the maintainance of cell shape. Tubulin itself is a globular protein which consists of two polypeptides (alpha and beta tubulin). Alpha and beta tubulin dimers are assembled to 13 protofilaments that form a microtubule of 22 nm diameter. Tyrosine ligase ads a C-terminal tyrosin 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. Alpha tubulin is not suitable as loading control in adipose tissue as expression of tubulin in adipose tissue is very low ( Spiegelman and Farmer, Cell, 1982, 29(1):53-60) in cells undergoing adipose differentiation actin synthesis decreases by 90%. Excellent as a protein loading control antibody.

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 4a Antibody (NB120-11304)(4)

Publications using NB120-11304

• Merritt EK, Stec MJ, Thalacker-Mercer A et al. Heightened muscle inflammation susceptibility may impair regenerative capacity in aging humans. J Appl Physiol (1985) 2013-09-01 [PMID: 23681911] (Human)
• Nicholas C. Mouse Polyomavirus T Antigens: Directors of Cell Cycle Signaling. Thesis. 2015-01-01 (WB, Mouse)
  
  Details:
  alpha Tubulin 4a antibody clone B-5-1-2 was used for WB analysis of lysates from C57 MEFs that were infected or not with MPyVs RA, NG59, and 808A (withan UI mock-infected control) for 1.5 hours. Tubulin immunoblots were used for normalization purpose for blots of NBS1, MRE11, RAD50, LTag / Large Tumor Antigen and LTAg (Figure 7).
• Ochaba J, Monteys AM, O'Rourke JG et al. PIAS1 Regulates Mutant Huntingtin Accumulation and Huntington's Disease-Associated Phenotypes In Vivo. Neuron. 2016-05-04 [PMID: 27146268] (WB, Mouse)
• Zaki MS, Bhat G, Sultan T et al. PYCR2 mutations cause a lethal syndrome of microcephaly and failure to thrive. Ann. Neurol. 2016-04-30 [PMID: 27130255] (WB, Human)

Bioinformatics

• Gene Symbol: TUBA4A
• Entrez:
  • 7277(Human)
  • 22143(Mouse)
  • 500929(Rat)