Customize nxf-1 Antibody

Description

Introduction to NXF-1 Antibody

The NXF-1 antibody is a polyclonal or monoclonal immunoglobulin designed to bind specifically to NXF-1, enabling its detection in assays such as Western blotting (WB), immunoprecipitation (IP), immunofluorescence (IF), and enzyme-linked immunosorbent assay (ELISA). NXF-1 shuttles between the nucleus and cytoplasm, facilitating the export of poly(A)+ RNA and retroviral RNA elements like the constitutive transport element (CTE) .

Key Features of NXF-1

Molecular Weight: ~70 kDa
Structure: Contains leucine-rich repeats (LRRs), an RNA-binding domain (RBD), and a ubiquitin-associated domain .
Function: Forms heterodimers with NXT1 (NTF2-related export protein 1) to interact with nucleoporins and mediate mRNA export .

Applications in Research

The NXF-1 antibody is instrumental in studying mRNA export mechanisms, viral pathogenesis, and cellular stress responses.

Common Applications

Application	Purpose	Key Studies
Western Blotting	Detect NXF-1 expression levels in cell lysates	Validated by Merck Millipore (ABE81) and Proteintech (10328-1-AP)
Immunoprecipitation	Identify NXF-1 protein complexes (e.g., TREX-2, nucleoporins)	Used in influenza virus studies to show NS1-BP-NXF1 interactions
Immunofluorescence	Localize NXF-1 to nuclear pore complexes or cytoplasmic stress granules	Demonstrated nuclear rim staining in TX-treated cells
ELISA	Quantify NXF-1 in biological samples	Validated by Proteintech (67914-1-Ig) and Abcepta (AP91997)

Role in Viral Pathogenesis

SARS-CoV-2: NXF-1 interacts with the viral Nsp1 protein to inhibit host mRNA export, suppressing antiviral responses. Antibodies targeting NXF1 revealed this interaction and identified critical residues (D33, E36, E37, E41) in Nsp1 .
Influenza Virus: NS1-BP protein binds NXF1 to mediate export of viral M mRNAs. Immunoprecipitation with NXF1 antibodies confirmed this interaction .
Retroviruses: NXF1 exports unspliced retroviral RNA (e.g., murine leukemia virus). Antibodies demonstrated NXF1 depletion reduces cytoplasmic viral RNA .

Cellular mRNA Export

TREX Complex: NXF1 associates with TREX components (e.g., Aly/REF, Thoc5) to license mRNA export. Antibodies showed that TREX disruption reduces NXF1 nuclear rim localization .
m6A-Modified mRNAs: NXF1 interacts with m6A-containing mRNAs via SRSF3 and YTHDC1. Antibodies validated this pathway in export studies .

Product Specs

Buffer

Preservative: 0.03% ProClin 300; Constituents: 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

14-16 week lead time (made-to-order)

Synonyms

nxf-1 antibody; C15H11.3 antibody; Nuclear RNA export factor 1 antibody

Target Names

nxf-1

Uniprot No.

Q9XVS7

Target Background

Function

Plays a critical role in the nuclear export of RNA to the cytoplasm.

Database Links

KEGG: cel:CELE_C15H11.3

STRING: 6239.C15H11.3a

UniGene: Cel.36637

Protein Families

NXF family

Subcellular Location

Nucleus. Note=Shuttle protein that accumulates in the nucleoplasm and at the nuclear rim.

Q&A

Here’s a structured FAQ collection for NXF1 antibody in academic research, incorporating methodological insights and data from peer-reviewed studies:

Advanced Research Questions

How to resolve contradictions in NXF1 localization data across studies?
- Case example: Nuclear vs. cytoplasmic roles in viral RNA export ( vs. ).
  - Experimental design adjustments:
    1. Use subcellular fractionation with markers (e.g., Lamin B1 for nucleus, GAPDH for cytoplasm) to confirm localization (as in ).
    2. Employ proximity ligation assays (PLA) to map interactions (e.g., NXF1-NS1-BP colocalization in A549 nuclei; ).
  - Critical controls:
    - Include siRNA-resistant NXF1 rescue constructs (e.g., 7.8-fold increase in nuclear NXF1 transcripts; ).
    - Use isotype-matched IgG and in situ negative controls (e.g., NS1-F103A/F138A mutant in ).
What multiplex strategies quantify NXF1 interactions with viral RNA export complexes?
- Integrated workflow:
  1. Co-immunoprecipitation (Co-IP): Use NXF1 antibody (1:50 dilution) with NXT1 or GANP antibodies to pull down TREX-2 complexes ( ).
  2. RNA-IP (RIP): Combine with qPCR for viral RNAs (e.g., MLV env or influenza NS1 mRNA; ).
  3. Live-cell imaging: Fuse NXF1 with GFP and track dynamics in infected 293T cells (see for granule localization protocols).
How to optimize NXF1 antibody for single-cell RNA export studies?
- Protocol refinement:
  - Spatial transcriptomics: Use IF-optimized NXF1 antibody (1:200) with RNA FISH probes targeting viral CTE elements ( ).
  - Single-molecule tracking: Pair with HaloTag-NXF1 constructs and Janelia Fluor ligands for real-time export kinetics ( ).

Data Interpretation Frameworks

Statistical thresholds for NXF1-dependent RNA export assays
- Benchmarks from literature ( ):
  
  Assay Significant Threshold Notes
  RT-qPCR (cytoplasmic RNA) ≥2-fold reduction post-KD P < 0.01 (Student’s t-test)
  PLA signal (interactions) ≥5 foci/cell Normalize to IgG controls
Troubleshooting low signal in NXF1 Co-IP experiments
- Solutions:
  - Increase crosslinker concentration (e.g., 2 mM DSP for nuclear complexes).
  - Pre-treat lysates with RNase A to disrupt RNA-mediated aggregates ( ).

Assay	Significant Threshold	Notes
RT-qPCR (cytoplasmic RNA)	≥2-fold reduction post-KD	P < 0.01 (Student’s t-test)
PLA signal (interactions)	≥5 foci/cell	Normalize to IgG controls

Emerging Applications

Role of NXF1 phosphorylation in mRNA granule formation
- Method: Use Phos-tag gels (WB) with λ-phosphatase-treated lysates to identify modifications affecting RNA binding (refer to ’s note on N-terminal phosphorylation sites).
NXF1 isoform-specific functions in neuronal cells
- Approach: Compare antibody reactivity against full-length NXF1 (70 kDa) vs. truncated isoforms (e.g., sNxf1 in polyribosomes; ).

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nxf-1 Antibody