Clone:
REA103
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MC
Alternative names:
CXCR5, BLR1, MDR15

Extended validation for CD185 (CXCR5) Antibody, anti-human, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA103
RF8B2++
MU5UBEE++
J252D4++
51505++
Cells were incubated with an excess of purified unconjugated CD185 (CXCR5) (REA103) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for CD185 (CXCR5). Human peripheral blood mononuclear cells (PBMCs) were stained with CD185 (CXCR5) antibodies and with a suitable counterstaining. As a control, CD185 (CXCR5) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD185 (CXCR5). Human peripheral blood mononuclear cells (PBMCs) were stained with CD185 (CXCR5) antibodies and with a suitable counterstaining. As a control, CD185 (CXCR5) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD185 (CXCR5). Human peripheral blood mononuclear cells (PBMCs) were stained with CD185 (CXCR5) antibodies and with a suitable counterstaining. As a control, CD185 (CXCR5) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD185 (CXCR5). Human peripheral blood mononuclear cells (PBMCs) were stained with CD185 (CXCR5) antibodies and with a suitable counterstaining. As a control, CD185 (CXCR5) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD185 (CXCR5). Human peripheral blood mononuclear cells (PBMCs) were stained with CD185 (CXCR5) antibodies and with a suitable counterstaining. As a control, CD185 (CXCR5) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for CD185 (CXCR5). Human peripheral blood mononuclear cells (PBMCs) were stained with CD185 (CXCR5) antibodies and with a suitable counterstaining. As a control, CD185 (CXCR5) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using CD185 (CXCR5) (REA103). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD185 (CXCR5) (REA103). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using CD185 (CXCR5) (REA103). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD185 (CXCR5) Antibody, anti-human, REAfinity™

Overview

Clone REA103 recognizes human CD185, which is a 42 kDa G-protein–coupled chemokine receptor and is also known as CXCR5, monocyte-derived receptor 15 (MDR15), or Burkitt lymphoma receptor-1 (BLR1). BLR-1 was originally identified on cells of Burkitt´s lymphoma and B cells. B cell attracting chemokine 1 (BCA-1), also known as B-lymphocyte chemoattractant (BLC) or CXCL13, is the ligand of CD185. It is further expressed on central memory CD4 T cells, T folicular helper cells, and on a migratory population of skin-derived dendritic cells.
Additional information: Clone REA103 displays negligible binding to Fc receptors.

Alternative names

CXCR5, BLR1, MDR15

Detailed product information

Technical specifications

CloneREA103
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD185 (CXCR5)
Alternative names of antigenCXCR5, BLR1, MDR15
Molecular mass of antigen [kDa]42
Distribution of antigenB cells, monocytes, neurons, T cells
Entrez Gene ID643
RRIDAB_2733205, AB_2811389, AB_2811386, AB_2811420, AB_2811408, AB_2819564, AB_2819538, AB_2801905, AB_2655783, AB_2655789, AB_2733204

References for CD185 (CXCR5) Antibody, anti-human, REAfinity™

Publications

  1. Bryant, V. L. et al. (2007)
    Cytokine-mediated regulation of human B cell differentiation into Ig-secreting cells: predominant role of IL-21 produced by CXCR5
    +
    T follicular helper cells.
    J. Immunol. 179: 8180-8190
  2. Chevalier, N. et al. (2011) CXCR5 expressing human central memory CD4 T cells and their relevance for humoral immune responses. J. Immunol. 186(10): 5556-5568
  3. Förster, R. et al. (1994) Expression of the G-protein-coupled receptor BLR1 defines mature, recirculating B cells and a subset of T-helper memory cells. Blood 84: 830-840
  4. Saeki, H. et al. (2000)
    A migratory population of skin-derived dendritic cells expresses CXCR5, responds to B lymphocyte chemoattractant
    in vitro
    , and co-localizes to B cell zones in lymph nodes
    in vivo
    .
    Eur. J. Immunol. 30(10): 2808-2814
  5. Förster, R. et al. (1996) A putative chemokine receptor, BLR1, directs B cell migration to defined lymphoid organs and specific anatomic compartments of the spleen. Cell 87(6): 1037-1047
  6. Farooq, F. et al. (2016) Circulating follicular T helper cells and cytokine profile in humans following vaccination with the rVSV-ZEBOV Ebola vaccine. Sci. Rep. 6: 27944
  7. García, M. et al. (2017) Peripheral T follicular helper cells make a difference in HIV reservoir size between elite controllers and patients on successful cART. Sci. Rep. 7(1): 16799

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