CD197 (CCR7) Antibody, anti-human, REAfinity™

Name: CD197 (CCR7) Antibody, anti-human, REAfinity™
Availability: InStock

CD197 (CCR7) Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA108

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MC

Alternative names:

CCR7, BLR-2, CC-CKR-7, CCR-7, Cdw197, CMKBR7, EBI-1

Flow cytometry applications forCD197 (CCR7) Antibody, anti-human, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 615
PE-Vio 770

Conjugate:

APC

Recommended dilution:

1:50

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Products used:

130-120-600, 130-120-460

Protocols:

Cell surface flow cytometry staining protocol for CCRs (1:50)

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD197 (CCR7) antibodies as well as with CD4 antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. Lymphocytes were gated for the analysis. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

Mass cytometry applications forCD197 (CCR7) Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Products used:

130-108-057

Extended validation for CD197 (CCR7) 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 clones	Overlap in epitope recognition with REA108
3D12	+
TG8	++
150503	++
G043H7	+
REA546	+
FR11-11E8	++
Cells were incubated with an excess of purified unconjugated CD197 (CCR7) (REA108) 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 CD197 (CCR7). Human peripheral blood mononuclear cells (PBMCs) were stained with CD197 (CCR7) antibodies and with a suitable counterstaining. As a control, CD197 (CCR7) 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 CD197 (CCR7). Human peripheral blood mononuclear cells (PBMCs) were stained with CD197 (CCR7) antibodies and with a suitable counterstaining. As a control, CD197 (CCR7) 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 CD197 (CCR7). Human peripheral blood mononuclear cells (PBMCs) were stained with CD197 (CCR7) antibodies and with a suitable counterstaining. As a control, CD197 (CCR7) 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 CD197 (CCR7). Human peripheral blood mononuclear cells (PBMCs) were stained with CD197 (CCR7) antibodies and with a suitable counterstaining. As a control, CD197 (CCR7) 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 CD197 (CCR7). Human peripheral blood mononuclear cells (PBMCs) were stained with CD197 (CCR7) antibodies and with a suitable counterstaining. As a control, CD197 (CCR7) 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 CD197 (CCR7). Human peripheral blood mononuclear cells (PBMCs) were stained with CD197 (CCR7) antibodies and with a suitable counterstaining. As a control, CD197 (CCR7) 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 CD197 (CCR7). Human peripheral blood mononuclear cells (PBMCs) were stained with CD197 (CCR7) antibodies and with a suitable counterstaining. As a control, CD197 (CCR7) 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 CD197 (CCR7) (REA108). 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 CD197 (CCR7) (REA108). 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 CD197 (CCR7) Antibody, anti-human, REAfinity™

Overview

REA108 recognizes human CD197 which is a chemokine receptor with a C-C motif that mediates homing of T cells to secondary lymphoid organs via high endothelial venules (HEV) and is also known as CCR7. For example, naive T cells migrate very efficiently through lymph nodes using the adhesion molecules L-selectin, CD62L, and CCR7. Ligands for CCR7, e.g. CCL19, are expressed by the HEV of secondary lymphoid organs, by parenchymal cells within T cell zones of lymph nodes, and by endothelial cells at the openings of lymphatic vessels within peripheral tissues. Expression of CCR7 and the CD45RA isoform distinguishes three subsets of T cells: naive T cells (CCR7

⁺

CD45RA

⁺

), central memory T cells (CCR7

⁺

CD45RA

^–

), and effector memory T cells (CCR7

^–

CD45RA

^–

).

Additional information: Clone REA108 displays negligible binding to Fc receptors.

Alternative names

CCR7, BLR-2, CC-CKR-7, CCR-7, Cdw197, CMKBR7, EBI-1

Detailed product information

Technical specifications

Clone	REA108
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (S), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human
Antigen	CD197 (CCR7)
Alternative names of antigen	CCR7, BLR-2, CC-CKR-7, CCR-7, Cdw197, CMKBR7, EBI-1
Molecular mass of antigen [kDa]	40
Distribution of antigen	B cells, dendritic cells, NK cells, macrophages, thymocytes
Entrez Gene ID	1236
RRID	AB_2751410, AB_2733212, AB_2733213, AB_2784046, AB_2784045, AB_2784044, AB_2784043, AB_2921736, AB_2921735, AB_2752158, AB_2752107, AB_2751042, AB_2904803, AB_2904802, AB_2751428

Resources for CD197 (CCR7) Antibody, anti-human, REAfinity™

Documents and Protocols

App notes/customer reports

Comprehensive flow cytometry analysis of chemokine receptors expressed on human natural killer cells.

Certificates

Please follow this

link

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

References for CD197 (CCR7) Antibody, anti-human, REAfinity™

Publications

Bacher, P. et al. (2014)
Identification of immunogenic antigens from
Aspergillus fumigatus
by direct multi parameter characterization of specific conventional and regulatory CD4
⁺
T cells.
J Immunol 193: 3332-3343
Sallusto, F. et al. (1999) Two subsets of memory T lymphocytes with distinct homing potentials and effector functions. Nature 401: 708-712
Baldan, W. et al. (2015) Efficient and reproducible generation of tumour-infiltrating lymphocytes for renal cell carcinoma. Br. J. Cancer 112(9): 1510-1518
Frascaroli, G. et al. (2018)
Human macrophages escape inhibition of major histocompatibility complex-dependent antigen presentation by
Cytomegalovirus
and drive proliferation and activation of memory CD4
⁺
and CD8
⁺
T cells.
Front Immunol 9
Tilly, G. et al. (2017) IL-15 Harnesses Pro-inflammatory Function of TEMRA CD8 in Kidney-Transplant Recipients. Front Immunol 8
Alvarez-Fernández, C. et al. (2016) A short CD3/CD28 costimulation combined with IL-21 enhance the generation of human memory stem T cells for adoptive immunotherapy. J. Transl. Med. 14: 214
Chang, Z. L. et al. (2015) Identification and selective expansion of functionally superior T cells expressing chimeric antigen receptors. J. Transl. Med. 13: 161

Applications

Extended validation