Product Information

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Product Background

Atypical chemokine receptor 1 (ACKR1), also known as Duffy antigen receptor for chemokines (DARC), is a seven-transmembrane atypical chemokine receptor involved in chemokine sequestration, inflammatory signaling regulation, leukocyte trafficking, vascular biology, and host-pathogen interactions.

ACKR1 is highly expressed in erythrocytes and endothelial cells and is widely studied in immunology, inflammation, infectious disease, hematology, vascular biology, and chemokine receptor signaling research.

Also known as: hACKR1, Human ACKR1, ACKR1, DARC, FY, CD234, Atypical chemokine receptor 1, Duffy antigen receptor for chemokines, Duffy antigen receptor, and Duffy blood group receptor.

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Antibody Development

HM0062 was generated by phage-display selection using a mouse-derived antibody library. Recombinant epitope-Fc fusion protein containing the ACKR1 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 human ACKR1 protein in human cell lines

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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 ACKR1 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 human ACKR1 protein in human cell lines under reducing conditions.

Validated samples:

• 293T cells
• HepG2 cells

Detection condition:

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

Observed bands:

• Higher molecular weight bands may represent protein glycosylation, oligomerization, or post-translationally modified species.