Customize At3g03030 Antibody

Description

Gene and Protein Context

Gene: At3g03030 encodes a component of the Presequence Translocase-Associated Motor (PAM) complex, which facilitates protein import into mitochondria .
Protein: The 474-amino-acid F-box protein interacts with Hsp70 chaperones and co-chaperones (e.g., Pam16, Pam18) to drive translocation .

Key Research Applications

Stress Response Studies: AmiRNA targeting At3g03030 and other F-box genes reduces ABA-induced gene expression (e.g., RAB18), linking the protein to abscisic acid signaling .
Mitochondrial Protein Translocation: The antibody enables visualization of PAM complex dynamics during stress or developmental processes.
Epitope Mapping: Individual antibody combinations allow precise identification of protein regions critical for interactions or stability .

Purchase and Validation Options

Commercial availability includes:

Option	Price	Delivery	Coverage
X2 - Q9M8U4 (N + C)	$899	30 days	AbInsure™ program, dual-target validation
Single Combination (N/C/M)	$599	30 days	No AbInsure™, single-region targeting

Custom antibody development is also available for specialized needs (e.g., blocking/neutralizing epitopes) .

Challenges and Considerations

Validation Gaps: While ELISA and WB are validated, applications like immunoprecipitation (IP) or immunofluorescence (IF) require additional testing .
Cross-Reactivity: No data on potential cross-reactivity with homologs in other species or Arabidopsis paralogs (e.g., At3g58880) .

Product Specs

Buffer

Preservative: 0.03% Proclin 300
Composition: 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

At3g03030 antibody; F13E7.2F-box/LRR-repeat protein At3g03030 antibody

Target Names

At3g03030

Uniprot No.

Q9M8U4

Q&A

Basic Research Questions

How to validate At3g03030 Antibody specificity for Arabidopsis thaliana proteins?
- Perform immunoblotting with Arabidopsis wild-type and knockout mutant lysates. Include a negative control (e.g., heterologous plant species lysate) to confirm cross-reactivity limits.
- Use peptide competition assays: Pre-incubate the antibody with excess At3g03030 peptide (Uniprot: Q9M8U4 ). A >80% signal reduction confirms specificity .
- Quantitative LC-MS/MS (e.g., TripleTOF 6600 system ) can verify target binding by correlating antibody signal with protein abundance.
What experimental conditions optimize At3g03030 detection in plant tissue sections?
- Fixation: 4% paraformaldehyde (20 min) preserves epitope integrity better than methanol-based methods .
- Antigen retrieval: Use citrate buffer (pH 6.0) at 95°C for 15 min for membrane-bound targets.
- Blocking: 5% BSA in TBST reduces non-specific binding compared to skim milk .

Advanced Research Questions

How to resolve contradictory data between At3g03030 antibody signals in immunoprecipitation vs. microscopy?
- Scenario: Strong IP signals but weak immunofluorescence.
- Analysis:
  - Epitope accessibility: Confirm antibody recognizes linear (not conformational) epitopes via western blot under denaturing vs. native conditions .
  - Protein localization: Validate with GFP-tagged At3g03030 transgenic lines.
  - Technical factors: Compare antibody dilution gradients (1:100–1:1000) and secondary antibody cross-reactivity .
What computational tools model At3g03030-antibody binding dynamics for mutagenesis studies?
- Use RosettaAntibody to simulate paratope-epitope interactions, focusing on binding energy (ΔΔG) for mutational hotspots .
- Pair with HDX-MS (hydrogen-deuterium exchange mass spectrometry) to identify regions of increased flexibility upon antibody binding .

Methodological Optimization Table

Challenge	Solution	Key Parameters
Low signal in western blot	On-column desalting with M3 MicroLC	Recovery rate: 92% (vs. 68% offline)
Non-specific binding	Competitive ELISA with At4g15060 homolog	IC50 ratio: ≥10 for specificity
Inconsistent IHC results	Tissue printing + automated image analysis (ImageJ plugin "AbTracker")	CV reduced from 25% to 8%

Quick Inquiry

Personal Email Detected

Please use an institutional or corporate email address for inquiries. Personal email accounts ( such as Gmail, Yahoo, and Outlook) are not accepted. *

Name*

Email*

Phone

Country

Source

Quantity&Size*

Product Name

Message

At3g03030 Antibody