Customize ABCG2 Antibody

Description

Introduction to ABCG2 Antibody

ABCG2 antibodies are immunological reagents designed to bind specifically to the ABCG2 protein, a transmembrane efflux transporter involved in drug resistance and cellular protection . These antibodies are used to study ABCG2's expression patterns, localization (e.g., plasma membrane, blood-brain barrier), and role in diseases such as cancer . Key clones include MAB995 (R&D Systems) and 5D3, which are widely cited in flow cytometry, immunohistochemistry (IHC), and immunocytochemistry (ICC) .

Applications in Research and Clinical Settings

ABCG2 antibodies are utilized for:

Flow Cytometry: Identifying ABCG2-expressing cells, such as hematopoietic stem cells and cancer stem cells .
Immunohistochemistry: Assessing ABCG2 expression in tumor tissues to predict chemotherapy resistance .
Drug Resistance Studies: Evaluating ABCG2-mediated efflux of chemotherapeutics like mitoxantrone and topotecan .

Validation and Specificity

Validation of ABCG2 antibodies is critical due to variability in performance across clones. A 2016 study tested six commercial antibodies, selecting one with high specificity for colorectal cancer (CRC) studies . Key validation steps include:

Western Blotting: Confirming protein size (~72 kDa monomer, 144 kDa dimer) .
Functional Assays: Measuring substrate transport inhibition (e.g., pheophorbide a efflux) .
Epitope Mapping: Ensuring binding to extracellular domains (e.g., 5D3 binds a conformational epitope) .

Role in Multidrug Resistance Studies

ABCG2 overexpression in cancers reduces intracellular drug accumulation, leading to treatment failure. Antibodies help correlate ABCG2 levels with clinical outcomes:

Breast Cancer: High ABCG2 expression correlates with poor pathological complete response (pCR) rates post-neoadjuvant chemotherapy .
Hepatocellular Carcinoma (HCC): Elevated ABCG2 predicts reduced survival in elderly patients .
Esophageal Cancer: ABCG2 knockdown via inhibitors (e.g., artesunate) reverses drug resistance .

Table 1: Clinical Correlations of ABCG2 Expression

Cancer Type	ABCG2 Expression	Clinical Outcome
Breast Cancer	High (78–80%)	Reduced pCR rate (19% vs. 38% low)
HCC	High	Poor survival in elderly patients
Colorectal Cancer	Variable	Linked to chemoresistance

Table 2: ABCG2 Antibody Performance

Clone	Application	Specificity	Key Use Case
MAB995	Flow Cytometry, ICC	High	Detects plasma membrane ABCG2
5D3	Conformational IHC	Moderate	Binds inhibitor-induced conformation
BXP-34	IHC (CRC validation)	High	Basolateral membrane staining in CRC

Challenges and Considerations

Specificity Issues: Non-specific binding observed with some clones (e.g., cytoplasmic vs. membrane staining) .
Intra-Tumor Heterogeneity: Variability in ABCG2 expression complicates biopsy interpretation .
Storage and Handling: Antibodies like MAB995 require optimized protocols for reproducible results .

Future Directions

Therapeutic Targeting: ABCG2 antibodies may guide inhibitor development (e.g., compound 8 in HCC) .
Biomarker Validation: Standardized scoring guidelines (similar to HER2) for clinical adoption .
Stem Cell Research: Exploring ABCG2's role in protecting stem cells from toxins .

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

ABCG2 antibody; WBC2 antibody; At2g37360 antibody; F3G5.15ABC transporter G family member 2 antibody; ABC transporter ABCG.2 antibody; AtABCG2 antibody; White-brown complex homolog protein 2 antibody; AtWBC2 antibody

Target Names

ABCG2

Uniprot No.

Q9ZUT0

Target Background

Database Links

KEGG: ath:AT2G37360

STRING: 3702.AT2G37360.1

UniGene: At.13323

Protein Families

ABC transporter superfamily, ABCG family, Eye pigment precursor importer (TC 3.A.1.204) subfamily

Subcellular Location

Membrane; Multi-pass membrane protein.

Q&A

ABCG2 antibodies are critical tools in cancer research, particularly for studying multidrug resistance mechanisms. Below is a structured FAQ collection addressing key methodological and analytical challenges in academic research:

What functional assays confirm ABCG2 antibody efficacy in modulating drug transport?

Advanced Research Focus

ATPase Activity Assay:
Measure ABCG2 ATP hydrolysis using crude membrane proteins (10 µg/100 µL) with beryllium fluoride inhibition .
- Key metric: Aurone-induced ATPase stimulation indicates direct interaction .
Radiolabeled Substrate Competition:
Use $^{125}\text{I}$ -IAAP photolabeling with test compounds (5 µM) to assess binding site competition .

Methodological Pitfall: Cytoplasmic staining ≠ functional activity. Prioritize membrane-localized ABCG2 in flow cytometry .

How to resolve contradictory ABCG2 expression data across studies?

Advanced Analytical Approach

Factor Analysis:

Variable	Impact on Data	Mitigation Strategy
Antibody Clone	Epitope specificity variations	Cross-validate with 2+ antibodies
Tissue Fixation	Epitope masking in FFPE	Antigen retrieval optimization
Cellular Conformation	IF vs. OF state detection	Use conformation-sensitive antibodies (e.g., 5D3)

Case Study: Discrepancies in Panc02.13 gemcitabine efflux data linked to Hippo-YAP pathway modulation of ABCG2 expression .

What structural insights guide antibody design for ABCG2 inhibition?

Advanced Mechanistic Research

Conformational Dynamics:
- Use 5D3 antibody to detect inward-facing (IF) vs. outward-facing (OF) states via flow cytometry .
- Substrate binding reduces 5D3 affinity during IF→OF transition ( $K_d$ shift from 2.1 nM → 15.4 nM) .
scFv Antibody Engineering:
- Phage display-derived scFv (e.g., anti-ABCG2 clones) reduces colony formation by 63% in lung adenocarcinoma via mitochondrial apoptosis .

How to select antibodies for live-cell vs. fixed-cell assays?

Basic Experimental Design

Live-Cell Requirements:
- Conformation-sensitive clones (e.g., 5D3 for IF state ).
- Non-internalizing epitopes (extracellular domain focus).
Fixed-Cell Validation:
- Prioritize antibodies recognizing linear epitopes resistant to formaldehyde cross-linking .

What in vivo models validate ABCG2 antibody therapeutic potential?

Advanced Translational Research

Xenograft Testing:
- MX-resistant nude mice: Aurones restore MX sensitivity (tumor volume reduction ≥40%) .
- Radioimmunoimaging: scFv antibodies show tumor-to-background ratios >3:1 at 72h post-injection .
Dosage Protocol:
- 10 mg/kg scFv biweekly + gemcitabine (100 mg/kg) synergizes apoptosis (↑ caspase-3 by 4.8-fold) .

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ABCG2 Antibody