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

Name: CD185 (CXCR5) Antibody, anti-mouse, REAfinity™
Availability: InStock

CD185 (CXCR5) Antibody, anti-mouse, REAfinity™Extended ValidationRecombinant

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

Flow cytometry applications forCD185 (CXCR5) Antibody, anti-mouse, REAfinity™

APC
Biotin
PE
PE-Vio 615
PE-Vio 770

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-119-129

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Spleen cells from C57/BL6 mice were stained with CD185 (CXCR5) antibodies as well as with CD45R (B220) antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. CD45.2⁺ cells were pre-gated for the analysis. The specificity of the conjugated antibodies was confirmed by blocking the binding to the ligand, using pure unconjugated antibodies (left images). Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

MICS (MACSima Imaging Cyclic Staining) applications forCD185 (CXCR5) Antibody, anti-mouse, REAfinity™

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

mouse

Products used:

130-119-129

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

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 clones	Overlap 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.

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.

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

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

Clone	REA215
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody, human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	mouse
Antigen	CD185 (CXCR5)
Alternative names of antigen	CXCR5, BLR1, CXC-R5, CXCR-5, Gpcr6, MDR15
Distribution of antigen	B cells, T cells
Entrez Gene ID	12145
RRID	AB_2733206, AB_2733207, AB_2727583, AB_2727601, AB_2802057, AB_2655794, AB_2655795, AB_2922210, AB_2751630

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

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

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

Publications

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
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

Applications

Extended validation