Clone:
REA215
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
CXCR5, BLR1, CXC-R5, CXCR-5, Gpcr6, MDR15

Extended validation for CD185 (CXCR5) Antibody, anti-mouse, 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 REA215
L138D7++
2G8++
SPRCL5++
Cells were incubated with an excess of purified unconjugated CD185 (CXCR5) (REA215) 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). Splenocytes from CD57BL/6 mice 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). Splenocytes from CD57BL/6 mice 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). Splenocytes from CD57BL/6 mice 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). Splenocytes from CD57BL/6 mice 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). Splenocytes from CD57BL/6 mice 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) (REA215). 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) (REA215). 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) (REA215). 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-mouse, REAfinity™

Overview

Clone REA215 recognizes murine CD185, which is a 42 kDa multipass chemokine receptor, also known as CXCR5 or burkitt lymphoma receptor-1 (BLR). Expression of CD185 is found on mature recirculating B cells, a small subset of CD4
+
and CD8
+
T cells, and skin-derived migratory dendritic cells. CXCR5 recognizes B cell–attracting chemokine 1 (BCA-1)/ CXCL13 as ligand, which is a homeostatic chemokine, constitutively secreted by stromal cells in B cell areas of secondary lymphoid tissues (follicles). CXCR5 is involved in recruitment of circulating naive B cells to follicles and trafficking and homing of B1 B cells. Mice lacking CXCR5 gene display defective primary follicles and germinal centers formation in the spleen and Peyer patches, and lack inguinal lymph nodes.
Additional information: Clone REA215 displays negligible binding to Fc receptors.

Alternative names

CXCR5, BLR1, CXC-R5, CXCR-5, Gpcr6, MDR15

Detailed product information

Technical specifications

CloneREA215
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD185 (CXCR5)
Alternative names of antigenCXCR5, BLR1, CXC-R5, CXCR-5, Gpcr6, MDR15
Distribution of antigenB cells, T cells
RRIDAB_2733206, AB_2733207, AB_2727583, AB_2727601, AB_2802057, AB_2655794, AB_2655795, AB_2751630

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

Publications

  1. Junt, T. et al. (2005) CXCR5-dependent seeding of follicular niches by B and Th cells augments antiviral B cell responses. J. Immunol. 175: 7109-7116
  2. Velaga, S. et al. (2009) Chemokine receptor CXCR5 supports solitary intestinal lymphoid tissue formation, B cell homing, and induction of intestinal IgA responses. J. Immunol. 182: 2610-2619

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