nxf-2 Antibody
Description
What is NXF-2 Antibody?
The NXF-2 antibody (e.g., Thermo Fisher 12185-1-AP) is a polyclonal antibody raised against a recombinant protein fragment derived from the human NXF2 sequence (amino acids 126–561) . It recognizes NXF2 across species, including humans, mice, and rats, and is validated for applications such as Western blotting (WB), immunohistochemistry (IHC), and immunofluorescence (IF) .
Development and Validation
-
Specificity: The antibody was validated using transfected cell lines expressing GFP-tagged NXF2 or untagged NXF2, with no cross-reactivity observed against NXF1 or unrelated proteins .
-
Immunohistochemistry: In testis and brain tissues, the antibody detected NXF2 in spermatogonia and hippocampal neurons, showing perinuclear and cytoplasmic localization .
Role in mRNA Dynamics and FMRP Interaction
-
NXF2 interacts with FMRP (Fragile X Mental Retardation Protein) in male germ cells and hippocampal neurons, forming RNA-binding complexes critical for mRNA stability and transport .
Cytoplasmic mRNA Localization
-
NXF2 binds motor proteins such as KIF17 and DyneinLC1, facilitating mRNA transport in neuronal dendrites .
Germ Cell-Specific Functions
-
In germ cells, NXF2 forms granular structures with XPO-1/CRM1 to anchor tra-2 mRNA at the nuclear periphery, preventing premature translation during meiosis .
Transposon Silencing
-
NXF2 partners with Piwi and Panx in Drosophila to silence transposons via piRNA-guided co-transcriptional repression .
Applications in Research
-
Western Blot: Detects NXF2 at ~80–100 kDa in transfected cells .
-
Immunofluorescence: Visualizes punctate cytoplasmic granules in neurons and germ cells .
-
Functional Studies: Used to explore NXF2’s roles in mRNA export, neuronal plasticity, and fertility .
Technical Considerations
Product Specs
Composition: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
Advanced Research Considerations
Why do NXF2-associated complexes exhibit cell-type-specific functional divergence?
-
Ovarian vs. testis contexts:
-
Experimental design: Perform compartment-specific IP (nuclear vs. cytoplasmic fractions) to map interaction networks (Fig. EV1E in ).
How to address variability in antibody-dependent NXF2 mechanistic studies?
-
Case example: Lab I and Lab II reported differing purity outcomes for in silico-generated antibodies (Fig. 7A in ):
Metric EXT Set (Clinical) GAN Set (In Silico) p-value Titer 120 ± 45 mg/L 150 ± 30 mg/L <0.05 Purity 85% ± 12% 92% ± 5% 0.08 -
Solutions:
What mechanisms underlie NXF2’s dual role in mRNA export and degradation?
-
Model: NXF2 recruits deadenylases/decapping enzymes to target mRNAs via its RRM domain.
-
Validation steps:
Technical Notes for Reproducibility
-
For WB, use 72 kDa as the expected molecular weight; deviations may indicate splice variants or post-translational modifications .
-
In IP, preclear lysates with Protein A/G beads to reduce nonspecific binding .
-
For epitope mapping, combine BLI-based competition assays with alanine scanning mutagenesis (Fig. 2a,e in ).
