Detection of Mouse and Rat DARC by Western Blot.
Western blot shows lysates of mouse liver tissue and rat liver tissue. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Mouse/Rat DARC Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6695) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for DARC at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of DARC in Mouse Splenocytes by Flow Cytometry.
Mouse splenocytes were stained with Rat Anti-Mouse TER‑119 APC‑conjugated Monoclonal Antibody (Catalog # FAB1125A) and either (A) Sheep Anti-Mouse/Rat DARC Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6695) or (B) Normal Sheep IgG Control (Catalog # 5-001-A) followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127).
Detection of DARC in HEK293 Human Cell Line Transfected with Mouse DARC and eGFP by Flow Cytometry.
HEK293 human embryonic kidney cell line transfected with mouse DARC and eGFP was stained with either (A) Sheep Anti-Mouse/Rat DARC Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6695) or (B) Normal Sheep IgG Control (Catalog # 5-001-A) followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127).
DARC in Mouse Skin.
DARC was detected in perfusion fixed frozen sections of mouse skin using Sheep Anti-Mouse/Rat DARC Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6695) at 0.5 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to the hair follicle. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
DARC (Duffy Antigen Receptor for Chemokines; also CD234) is a 40-46 kDa glycoprotein member of the Duffy family of silent heptahelical chemokine receptors. It is expressed in liver and on select neurons, erythrocytes and the endothelium of postcapillary venules. Unlike traditional chemokine receptors, DARC cannot signal through G-proteins as it lacks a DRYLAIVHA cytoplasmic motif. DARC has three potential functions: first, it binds circulating inflammatory-type chemokines, serving as a repository for future chemokine release; second, it acts as a vehicle by which chemokines are transported from the abluminal to the luminal side of endothelium; and third, it complexes with signal-transducing chemokine receptors to create a nonsignaling heterodimer. Mouse DARC is 334 amino acids (aa) in length. It contains a 62 aa N-terminal extracellular region, and a 28 aa C-terminal cytoplasmic tail. There is one potential splice variant that shows a 42 aa substitution for aa 133-334. Collectively, over the four extracellular domains (aa 1-62, 115-127, 186-205, 264-285), mouse DARC shares 52% and 75% aa identity with human and rat DARC, respectively.
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