TUBE1 Monoclonal Antibody

What is TUBE1 and what cellular functions does it serve?

TUBE1, also known as Epsilon-tubulin or Tubulin epsilon chain, is a member of the tubulin protein family involved in cellular structure and microtubule organization . Unlike the more extensively characterized alpha and beta tubulins, epsilon tubulin has distinct functions in microtubule nucleation and organization. The protein has a calculated molecular weight of approximately 52,932 Da according to technical specifications .

What applications are TUBE1 monoclonal antibodies validated for?

Current research-grade TUBE1 monoclonal antibodies have been validated for multiple applications including:

Each antibody should be individually optimized for specific experimental conditions to ensure optimal results .

What species reactivity can be expected from available TUBE1 antibodies?

Most commercially available TUBE1 monoclonal antibodies demonstrate reactivity against human, mouse, and rat species . For example, the monoclonal antibody with clone number 2B3 (IgG isotype) shows confirmed reactivity to TUBE1 across these three species .

What are the critical parameters for successfully using TUBE1 antibodies in Western blotting?

For optimal Western blotting results with TUBE1 monoclonal antibodies:

• Recommended dilution ranges typically fall between 1:1000-1:2000 or 1:500-1:5000 depending on the specific antibody clone

• Validation data indicates successful detection in MCF7 cell lysates as a positive control

• The expected band size corresponds to the calculated molecular weight of approximately 53 kDa

• Standard Western blotting protocols apply, with attention to proper blocking and washing steps

Researchers should verify specific band detection before proceeding with experimental analyses.

How should researchers validate TUBE1 antibodies prior to experimental use?

Proper validation should include:

• Specificity testing: Verification using known positive samples (e.g., MCF7 cells for certain antibodies)

• Signal verification: Confirmation of appropriate molecular weight bands in Western blot applications

• Cross-reactivity assessment: Testing against potential cross-reactive proteins, particularly other tubulin family members

• Immunogen verification: Understanding the specific peptide sequence used for antibody generation (synthetic peptides of Epsilon Tubulin are common immunogens)

• Batch testing: Verifying consistency between different lots of the same antibody

What storage conditions maintain optimal TUBE1 antibody activity?

Most TUBE1 monoclonal antibodies require:

• Storage in buffers containing stabilizers (typically PBS with 50% glycerol, 0.5% BSA, and 0.02% sodium azide)

• Long-term storage at recommended temperatures (typically -20°C)

• Minimal freeze-thaw cycles to preserve antibody integrity

• Protection from contamination and degradation

How can recombinant antibody technology be applied to TUBE1 research?

Recent advances in recombinant antibody technology offer significant advantages for TUBE1 research:

• Production methods can generate high-yield recombinant monoclonal antibodies from human HEK293 suspension cultures, addressing concerns with reproducibility, costs, and ethical issues associated with animal-derived antibodies

• The protocol involves:

  • Generating antibody heavy and light chain plasmids from a primary sequence

  • Transfecting plasmids into cells

  • Purifying antibodies using cost-effective methods

• This approach yields consistent antibodies with potentially increased experimental flexibility

• Researchers can introduce sequence modifications to enhance specificity for TUBE1 over other tubulin family members

What purification methods yield the highest quality TUBE1 monoclonal antibodies?

Optimal purification methodologies include:

• Affinity chromatography: TUBE1 antibodies can be effectively purified from mouse ascites or cell culture supernatants using affinity chromatography with specific immunogens

• Column-based techniques: These provide high purity while maintaining antibody activity

• Low-cost approaches: As described in recent literature, cost-effective purification methods can produce high-yield reagents suitable for research applications

How does TUBE1 antibody selection differ from other tubulin family antibodies?

When selecting between tubulin family antibodies:

• Alpha and beta tubulin antibodies (such as Anti-β-Tubulin mouse monoclonal antibodies) are more commonly used for general cytoskeletal studies

• TUBE1 antibodies target epsilon tubulin specifically and serve more specialized research purposes

• Many researchers using tubulin antibodies also investigate related products such as:

  • Anti-β-Tubulin III rabbit antibodies

  • Anti-Tubulin rabbit antibodies

  • Anti-α-Tubulin mouse antibodies

What unique epitopes distinguish TUBE1 monoclonal antibodies from other tubulin antibodies?

TUBE1 monoclonal antibodies typically:

• Target synthetic peptide sequences specific to Epsilon Tubulin

• Require careful epitope selection to avoid cross-reactivity with other tubulin family members

• May utilize specific regions of the protein that differ from the highly conserved domains shared across the tubulin family

How are monoclonal antibodies being utilized in pain-related research?

While not specific to TUBE1, monoclonal antibody research is advancing in chronic pain treatment:

• Monoclonal antibodies target specific molecules involved in pain signaling pathways, offering novel therapeutic approaches

• Current targets include tumor necrosis factor (TNF), nerve growth factor (NGF), calcitonin gene-related peptide (CGRP), and interleukin-6 (IL-6)

• These approaches are being investigated for conditions including osteoarthritis, chronic lower back pain, migraine, and rheumatoid arthritis

• Preclinical research suggests potential for additional targets in pain processing pathways

What methodological considerations are important when using monoclonal antibodies for epitope binding studies?

Recent structural analysis of neutralizing monoclonal antibodies provides insights applicable to TUBE1 research:

• Binding characteristics can be determined through site-directed alanine mutations of potential epitope residues

• ELISA testing of mutant antigens can identify key contact residues

• Complementarity determining regions (CDRs) of antibody heavy and light chains interact with specific protein loops to create binding footprints

• Buried surface area measurements provide quantitative data on antibody-antigen interaction strength

How should researchers address non-specific binding issues with TUBE1 antibodies?

To minimize non-specific binding:

• Optimize blocking conditions using appropriate blocking agents (BSA, non-fat milk, or commercial blockers)

• Titrate antibody concentrations to determine optimal signal-to-noise ratios

• Increase washing stringency in immunoassay protocols

• Consider using monoclonal antibodies with more defined epitope specificity

• Implement negative controls lacking primary antibody to identify sources of background

What controls are essential when validating experimental results with TUBE1 antibodies?

Essential controls include:

• Positive controls: Cell lines known to express TUBE1 (e.g., MCF7 cells for some antibody clones)

• Negative controls: Samples known to have minimal TUBE1 expression or knockout/knockdown models

• Isotype controls: Matched isotype antibodies to control for non-specific binding

• Secondary antibody-only controls: To identify background from detection systems

• Peptide competition assays: Using immunizing peptides to confirm specificity