At2g23060 Antibody

The At2g23060 antibody is a polyclonal reagent targeting a protein encoded by locus AT2G23060 in Arabidopsis thaliana, widely used in plant molecular biology research. Below are structured FAQs addressing key methodological considerations for academic researchers, derived from technical specifications and research applications .

Advanced Methodological Considerations

How to resolve non-specific bands in Western blots using this antibody?

• Implement tiered blocking:

  • 5% non-fat milk in TBST (1 hr)

  • Primary antibody incubation at 4°C overnight with 0.1% Tween-20

  • Secondary antibody cross-adsorbed against Arabidopsis proteome

What experimental controls are critical when studying At2g23060 protein-protein interactions?

How to adapt ELISA protocols for quantitative analysis of At2g23060 expression?

• Standard curve: 0.1-100 ng/mL recombinant protein (R² ≥ 0.98)

• Plate coating: 2.5 μg/mL capture antibody in carbonate buffer (pH9.6)

• Signal amplification: Biotin-streptavidin-HRP system with TMB substrate

Technical Troubleshooting

What are the implications of batch-to-batch variability in polyclonal antibodies?

• Maintain detailed records of:

  • Immunogen lot (recombinant protein sequence alignment)

  • ELISA titer values across experimental batches

  • Cross-reactivity profiles with Arabidopsis homologs

How to validate antibody performance in mutant complementation lines?

• Confirm genomic insertion via PCR (35 cycles, Tm 58°C)

• Western blot with 15% SDS-PAGE and 2% SDS extraction buffer

• Quantify signal intensity against actin control (ImageJ v1.53)