Customize EZA1 Antibody

Description

Definition and Structure of EEA1 Antibody

The EEA1 Antibody (e.g., ab2900 from Abcam) is a rabbit polyclonal antibody targeting Early Endosome Antigen 1 (EEA1), a zinc finger FYVE domain-containing protein critical for early endosomal membrane trafficking . EEA1 binds phosphatidylinositol 3-phosphate (PI3P)-enriched vesicles and facilitates endosomal fusion through interactions with Rab5 GTPases .

Characteristic	Detail
Antigen	EEA1 (ZFYVE2)
Format	Polyclonal rabbit antibody
Applications	Western blot (WB), immunocytochemistry (ICC/IF)
Species Compatibility	Human, mouse, rat, and others
Predicted Band Size	162 kDa
Observed Bands	100 kDa, 180 kDa, 41 kDa, 50 kDa (cross-reactive)

Western Blot Analysis

The EEA1 Antibody is validated for detecting EEA1 in lysates from human cell lines (e.g., HeLa, A431, Jurkat) and primary cultures (e.g., rat hippocampal neurons) . Key findings include:

HeLa nuclear lysate: Bands at 100 kDa and 180 kDa .
Whole-cell lysates: Additional bands at 41 kDa and 50 kDa observed, suggesting cross-reactivity or degradation products .
Knockout validation: Signal loss in EEA1 knockout HAP1 cells confirms specificity .

Immunocytochemistry

The antibody enables visualization of early endosomes in fixed and permeabilized cells. For example:

HepG2 cells: Strong punctate staining in the cytoplasm, colocalizing with α-tubulin .
Primary rat neurons: EEA1 colocalizes with β-tubulin in dendritic processes, indicating endosomal trafficking dynamics .

Endosomal Trafficking

EEA1 serves as a marker for early endosomes, which function in sorting internalized receptors and pathogens. Studies using this antibody have revealed:

Membrane remodeling: EEA1 facilitates homotypic fusion of early endosomes through PI3P recognition .
Disease relevance: Dysregulation of EEA1 is implicated in neurodegenerative disorders (e.g., Alzheimer’s disease) and viral entry mechanisms .

Cross-reactivity and Limitations

While the EEA1 Antibody is highly specific, non-target bands (41 kDa, 50 kDa) are observed in some samples, potentially reflecting post-translational modifications or epitope sharing .

The EEA1 Antibody is indispensable for studying:

Membrane trafficking pathways: Elucidating mechanisms of receptor recycling and pathogen entry.
Neurological diseases: Investigating endosomal dysfunction in Alzheimer’s and Parkinson’s.
Viral replication: Mapping early endosomal stages in viral life cycles (e.g., HIV, SARS-CoV-2) .

Product Specs

Buffer

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

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

EZA1 antibody; SDG10 antibody; SET10 antibody; SWN antibody; At4g02020 antibody; T10M13.3Histone-lysine N-methyltransferase EZA1 antibody; EC 2.1.1.356 antibody; CURLY LEAF-like 1 antibody; Protein SET DOMAIN GROUP 10 antibody; Protein SWINGER antibody

Target Names

EZA1

Uniprot No.

Q9ZSM8

Target Background

Function

EZA1 is a Polycomb group (PcG) protein that serves as the catalytic subunit of certain PcG multiprotein complexes. These complexes are responsible for methylating 'Lys-27' of histone H3, leading to transcriptional repression of the affected target genes. PcG proteins function by forming multiprotein complexes, which are essential for maintaining the transcriptionally repressive state of homeotic genes throughout development. While PcG proteins are not involved in initiating repression, they play a crucial role in sustaining this repression during later developmental stages.

Gene References Into Functions

Expression of embryo development genes is repressed by PRC1-catalytic core subunits AtBMI1 and AtRING1, a mechanism shared with PRC2-catalytic enzymes CLF or SWINGER (SWN). PMID: 26760036
MEA and SWN, as SET-domain partners of FIS2, perform partially redundant functions in regulating the initiation of endosperm development prior to fertilization in Arabidopsis. [SWINGER] [SWN] PMID: 16924116

Database Links

KEGG: ath:AT4G02020

STRING: 3702.AT4G02020.1

UniGene: At.3858

Protein Families

Class V-like SAM-binding methyltransferase superfamily, Histone-lysine methyltransferase family, EZ subfamily

Subcellular Location

Nucleus.

Q&A

Here’s a structured FAQ collection for "EZA1 Antibody" tailored to academic research, synthesized from peer-reviewed methodologies and engineering principles in the provided sources:

Advanced Research Questions

How to resolve discrepancies in EZA1 binding affinity across assay platforms?

Troubleshooting steps:
- Quantify avidity effects (e.g., compare monovalent Fab vs. bivalent IgG formats) .
- Assess antigen density (e.g., flow cytometry vs. ELISA may report different K<sub>D</sub> due to valency) .
- Control for assay-specific variables (e.g., pH, ionic strength in SPR vs. cell-based assays) .

What engineering strategies improve EZA1’s manufacturability without compromising function?

Approaches:

Issue	Solution	Outcome
Low expression	Humanize VH/VL frameworks (e.g., Prometheus™ platform)	30x higher titer
Aggregation	Switch to favorable Fc subtypes (e.g., IgG2/4)	Monomer content >95%
Instability	Glyco-engineering (e.g., afucosylation for improved FcγR binding)

How to optimize EZA1’s Fc effector function for immuno-oncology applications?

Design considerations:
- Pro-inflammatory: Use IgG1 with FcγR-enhancing mutations (e.g., GASDALIE) .
- Anti-inflammatory: Employ Fc Silent™ mutations (e.g., L234A/L235A) to abolish ADCC/CDC .
- Validate using NF-κB reporter assays for FcγR signaling .

How to integrate EZA1 into an ADC (Antibody-Drug Conjugate) workflow?

Critical parameters:
- Maintain Drug-Antibody Ratio (DAR) between 3.4–4.4 via Design of Experiments (DoE) .
- Monitor conjugate stability under stress conditions (e.g., pH 5.0 for lysosomal mimicry) .
- Use orthogonal analytics (SEC, HIC, CE-SDS) to assess aggregation and payload distribution .

Data Contradiction Analysis

How to address conflicting results in EZA1’s target engagement across studies?

Root-cause analysis:
- Compare antibody batches (track lot-specific glycosylation profiles) .
- Re-evaluate epitope accessibility (e.g., use truncated antigen variants) .
- Standardize assay conditions (e.g., fixatives in IHC alter epitope conformation) .

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

EZA1 Antibody