Product Information

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Product Background

Retinoic acid receptor gamma (RARγ / RARG) is a ligand-dependent nuclear receptor involved in retinoic acid signaling, transcriptional regulation, cellular differentiation, embryonic development, stem cell biology, and cancer-related signaling pathways. RARγ is widely studied in developmental biology, oncology, stem cell regulation, and nuclear receptor signaling research.

RARγ functions as a transcription factor that regulates retinoid-responsive genes and plays important roles in tissue differentiation and cellular homeostasis.

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Antibody Development

XM0044 was generated by phage-display selection using a mouse-derived antibody library. Recombinant epitope-Fc fusion protein containing the RARγ epitope sequence was used as the selection antigen. Following identification of the initial antibody candidate, structure-guided affinity optimization was performed to improve binding affinity and specificity.

The optimized antibody was reconstructed as a full-length mouse IgG1 recombinant antibody and expressed in HEK293 cells.

Binding and specificity were evaluated by:

• ELISA using recombinant epitope-Fc fusion antigen
• Western blot using recombinant MBP-epitope fusion protein
• Western blot detection of endogenous RARγ protein in human cells and mouse tissue

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Validation Data

ELISA Validation

ELISA demonstrated specific binding to recombinant epitope-Fc fusion antigen, while minimal binding was observed against Fc control protein.

Validation conditions:

• Well 1: Fc control protein
• Well 2: Epitope-Fc fusion protein
• Detection substrate: TMB

Note:
ELISA validation confirms binding to the epitope-Fc fusion antigen. Full-length RARγ protein binding in solution has not been tested by ELISA.

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Western Blot Validation – Recombinant Fusion Protein

Western blot analysis demonstrated specific recognition of recombinant MBP-epitope fusion protein expressed in BL21(DE3) cells, while no significant binding was observed against MBP control protein.

Validation conditions:

• Lane 1: MBP control protein
• Lane 2: MBP-epitope fusion protein
• Standard reducing Western blot

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Western Blot Validation – Endogenous Full-Length Protein

Western blot analysis detected endogenous RARγ protein in HeLa cells and mouse heart tissue under reducing conditions.

Validated samples:

• HeLa cells
• Mouse heart tissue

Detection condition:

• Standard reducing Western blot
• HRP-conjugated anti-mouse secondary antibody

Observed bands:

• Observed bands located at around 40Kd.