Customize FIP1 Antibody

Description

FIP1 vs. FIP1L1: Clarifying the Context

While FIP1 is a subunit of the CPSF complex critical for polyadenylation (mRNA 3′-end formation), FIP1L1 is a separate gene product known for its role in clonal hypereosinophilic syndromes due to its fusion with PDGFRA. Antibodies targeting these proteins serve distinct purposes:

FIP1 Antibodies: Focus on studying mRNA processing mechanisms.
FIP1L1 Antibodies: Primarily used to diagnose FIP1L1-PDGFRA fusion-related diseases.

Current literature emphasizes FIP1L1 antibodies in clinical diagnostics, while FIP1-specific tools remain less documented. This distinction is critical for interpreting antibody applications.

Diagnostic Use in Hypereosinophilic Syndromes

FIP1L1 antibodies are pivotal in detecting the FIP1L1-PDGFRA fusion, a hallmark of clonal hypereosinophilic syndromes (HES). Key findings:

Imatinib Responsiveness: Patients with FIP1L1-PDGFRA fusion exhibit dramatic responses to imatinib, a tyrosine kinase inhibitor .
Stroke Association: FIP1L1-PDGFRA+ HES is linked to ischemic strokes with watershed infarctions, often undetermined by TOAST criteria .

Mechanistic Insights

FIP1L1 antibodies aid in studying its role beyond HES:

mRNA Processing: FIP1 (not FIP1L1) binds U-rich RNA elements via its arginine-rich domain, facilitating poly(A) site recognition .
Kidney Injury: FIP1 (distinct from FIP1L1) exacerbates kidney damage by inducing NLRP3 3′UTR shortening, amplifying inflammation .

FIP1 Antibody Limitations and Future Directions

While FIP1L1 antibodies are well-characterized, FIP1-specific tools remain underexplored. Gaps include:

CPSF Complex Studies: Limited data on FIP1’s structural interactions (e.g., with CPSF30, PAP) in human systems .
Therapeutic Potential: FIP1’s role in alternative polyadenylation (APA) regulation in stem cells or cancer warrants further antibody-based exploration .

Product Specs

Buffer

Preservative: 0.03% Proclin 300
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

FIP1 antibody; At1g28200 antibody; F3H9.14GEM-like protein 1 antibody; Forming homology-interacting protein 1 antibody; FH-interacting protein 1 antibody

Target Names

FIP1

Uniprot No.

Q9SE96

Q&A

FAQs for FIP1 Antibody Research Applications

Advanced Research Questions

How to resolve discrepancies in FIP1 molecular weight across studies?

Critical analysis:
- Observed: 67 kDa in WB vs. calculated: 66 kDa (594 aa) . This 1 kDa difference may arise from post-translational modifications (e.g., phosphorylation) .
- Troubleshooting:
  - Compare lysate preparation methods (RIPA vs. NP-40 buffers).
  - Validate with recombinant FIP1 protein (1:1 binding to poly(A) polymerase) .

How to design co-immunoprecipitation (Co-IP) experiments for FIP1 interactome mapping?

Protocol:
- Crosslink ESCs with formaldehyde to preserve transient RNA-protein interactions .
- Use anti-FIP1 antibody (≥1:500 dilution) with Protein A/G beads.
- Validate interactions via Western blot for known partners (e.g., RNA14 or CPSF subunits) .
- Key finding: FIP1 binds U-rich sequences upstream of AAUAAA motifs (iCLIP-seq data) .

How to address conflicting APA regulation data in FIP1-depleted systems?

Case example:
- ESC vs. MEFs: FIP1 KD in ESCs causes significant PtoD APA shifts (311 genes) , but minimal APA changes in MEFs .
- Resolution:
  - Quantify 3′ UTR isoforms using DRS or 3′-RACE .
  - Control for cell-type-specific CPSF complex composition (e.g., RNA14 expression) .

Technical Challenges & Solutions

Why does FIP1 antibody exhibit non-specific bands in WB?

Strategies:
- Pre-adsorb antibody with FIP1L1 fusion protein .
- Compare signal intensity in WT vs. FIP1-KO cells (e.g., A431 Rab11-FIP1-KO) .

How to link FIP1-mediated APA to functional phenotypes (e.g., pluripotency)?

Integrated workflow:
- Perform RNA-seq + APA analysis post-FIP1 KD .
- Cross-reference with proteomics data to identify shortened 3′ UTR-associated proteins (e.g., Cdkn2b) .
- Validate using luciferase reporters with proximal/distal PASs .

Data Interpretation Tables

Table 1. FIP1 antibody validation across species ( )

Species	WB	IHC	IF/ICC
Human	Yes	Yes	Yes
Mouse	Yes	No data	No data
Rat	Yes	No data	No data

Table 2. FIP1 functional roles in APA regulation ( )

Function	Mechanism	Key Evidence
Proximal PAS preference	Binds U-rich elements near AAUAAA motifs	iCLIP-seq (P = 1.1 × 10⁻¹⁸)
Poly(A) polymerase tethering	Forms 1:1 complex with PAP1	Yeast two-hybrid + Co-IP
Reprogramming efficiency	Regulates early iPSC marker induction	88-fold reduction in Oct4-EGFP+ cells

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FIP1 Antibody