KLRG1 Antibody, anti-human, REAfinity™

Name: KLRG1 Antibody, anti-human, REAfinity™
Availability: InStock

KLRG1 Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA261

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC, MC

Alternative names:

Killer cell lectin-like receptor subfamily G member 1, CLEC15A, MAFA, MAFA-2F1, MAFA-L, 2F1

Flow cytometry applications forKLRG1 Antibody, anti-human, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700
Vio Bright B515
VioBlue

Conjugate:

APC

Recommended dilution:

1:50

Remark:

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

Tested:

QC tested

Products used:

130-117-815, 130-117-703

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with Anti-KLRG1 antibodies as well as with CD56 antibodies. The specificity of the conjugated antibodies is confirmed by blocking the binding to ligand, using pure unconjugated antibodies (left images). Cells were then analyzed by flow cytometry using the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. 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.

MICS (MACSima Imaging Cyclic Staining) applications forKLRG1 Antibody, anti-human, REAfinity™

PE

Conjugate:

PE

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-120-566, 130-120-426

Tissue used:

tonsil

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Description:

Immunofluorescence analysis of a human tonsil cryosection fixed with ice-cold acetone. The section was stained using KLRG1 Antibody, anti-human, PE (green) for 10 min at room temperature.

Mass cytometry applications forKLRG1 Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Products used:

130-126-458

Extended validation for KLRG1 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 REA261
13A2	++
13F12F2	++
SA231A2	++
14C2A07	+
Cells were incubated with an excess of purified unconjugated KLRG1 (REA261) 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 KLGR1. Human peripheral blood mononuclear cells (PBMCs) were stained with KLGR1 antibodies and with a suitable counterstaining. As a control, KLGR1 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 KLGR1. Human peripheral blood mononuclear cells (PBMCs) were stained with KLGR1 antibodies and with a suitable counterstaining. As a control, KLGR1 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 KLGR1. Human peripheral blood mononuclear cells (PBMCs) were stained with KLGR1 antibodies and with a suitable counterstaining. As a control, KLGR1 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 KLGR1. Human peripheral blood mononuclear cells (PBMCs) were stained with KLGR1 antibodies and with a suitable counterstaining. As a control, KLGR1 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 KLGR1. Human peripheral blood mononuclear cells (PBMCs) were stained with KLGR1 antibodies and with a suitable counterstaining. As a control, KLGR1 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 KLRG1 (REA261). 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 KLRG1 (REA261). 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 KLRG1 Antibody, anti-human, REAfinity™

Overview

Clone REA261 recognizes the killer cell lectin-like receptor subfamily G member 1 (KLRG1) antigen, a transmembrane protein, which is also known as C-type lectin domain family 15 member A (CLEC15A) or ITIM-containing receptor MAFA-L. KLRG1 is expressed by basophils, CD4 and CD8 T cells that exhibit a memory cell phenotype, and by a large proportion of peripheral blood NK cells. Expression of KLRG1 by virus-specific CD8

⁺

T cells is induced by repetitive antigen stimulation and it differs in chronic versus resolved viral infections. T cells expressing KLRG1 exhibit a poor proliferation potential, while their ability to secrete gamma interferon is preserved. Thus, KLRG1 is a convenient marker for early T cell–differentiation.

Additional information: Clone REA261 displays negligible binding to Fc receptors.

Alternative names

Killer cell lectin-like receptor subfamily G member 1, CLEC15A, MAFA, MAFA-2F1, MAFA-L, 2F1

Detailed product information

Technical specifications

Clone	REA261
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	KLRG1
Alternative names of antigen	Killer cell lectin-like receptor subfamily G member 1, CLEC15A, MAFA, MAFA-2F1, MAFA-L, 2F1
Molecular mass of antigen [kDa]	22
Distribution of antigen	T cells, NK cells, basophils
Entrez Gene ID	10219
RRID	AB_2752143, AB_2752091, AB_2784406, AB_2784405, AB_2784408, AB_2784407, AB_2784410, AB_2784409, AB_2784404, AB_2784403, AB_2752142, AB_2752090, AB_2811538, AB_2811530, AB_2889702, AB_2889704, AB_2889703, AB_2733432, AB_2733431

Resources for KLRG1 Antibody, anti-human, REAfinity™

Certificates

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References for KLRG1 Antibody, anti-human, REAfinity™

Publications

Voehringer, D. et al. (2002) Lack of proliferative capacity of human effector and memory T cells expressing killer cell lectinlike receptor G1 (KLRG1). Blood 100: 3698-3702
Butcher, S. et al. (1998) MAFA-L, an ITIM-containing receptor encoded by the human NK cell gene complex and expressed by basophils and NK cells. Eur. J. Immunol. 28(11): 3755-3762
Thimme, R. et al. (2005) Increased expression of the NK cell receptor KLRG1 by virus-specific CD8 T cells during persistent antigen stimulation. J. Virol. 79: 12112-12116

Applications

Extended validation