Description
The 13E11 monoclonal antibody specifically binds to CC chemokine receptor-like 1 (CCRL1) that is also known as C-C chemokine receptor type 11 (C-C CKR-11), or ChemoCentryx chemokine receptor (CCX-CKR). CCRL1 is a seven-transmembrane glycoprotein receptor that is encoded by ACKR4 (Atypical chemokine receptor 4) which belongs to the atypical chemokine receptor subfamily within the chemokine receptor superfamily. CCRL1 is expressed on stromal cells of lymph nodes, dermal lymphatic endothelial cells, and thymic epithelial cells. CCRL1 binds to several "homeostatic" chemokines including CCL19 (ELC), CCL21 (SLC), CCL25 (TECK) and CXCL13 (BLC) that can otherwise bind to functional chemokine receptors like CCR7, CCR9 or CXCR5. CCRL1 can thus serve as a decoy receptor that allows cellular uptake and degradation of these chemokines in a G protein-independent manner. This receptor may shape chemokine gradients in tissues by scavenging chemokines and thereby regulate the chemotaxis of thymic precursor cells, various leucocytes, including lymphocytes and dendritic cells, and cancer cells. CCRL1 expression is upregulated on some breast and hepatic cancer cell lines. The 13E11 antibody reportedly has ligand-like activity that results in the cellular internalization of CCRL1. This antibody recognizes an epitope in the N-terminus of the first extracellular domain of CCRL1.
Bead-based compensation or unmixing controls, such as BD® CompBeads or BD™ SpectraComp™, can be used as surrogates to assess fluorescence spillover when bound to fluorochrome-conjugated antibodies. Although these beads have spectral properties similar to cells, variations in spectral emission may occur, resulting in differing spillover values compared to biological controls. Therefore, it is considered best practice to compare the spillover obtained from cells and bead-based controls when using BD® CompBeads or BD™ SpectraComp™ for the first time, to ensure they are appropriate for the intended application.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.
1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
3. For U.S. patents that may apply, see bd.com/patents.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
6. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
7. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
9. An isotype control should be used at the same concentration as the antibody of interest.
10. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
11. Alexa Fluor™ is a trademark of Life Technologies Corporation.
12. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
