CD271 (LNGFR) Antibody, anti-human, REAfinity™

Name: CD271 (LNGFR) Antibody, anti-human, REAfinity™
Availability: InStock

CD271 (LNGFR) Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA844

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC

Alternative names:

NGFR, Gp80-LNGFR, TNFRSF16, p75(NTR), p75NTR, p75NGFR, NGFR (p75)

Flow cytometry applications forCD271 (LNGFR) Antibody, anti-human, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 615
PE-Vio 770
Vio Bright B515
Vio Bright R720
VioBlue
VioGreen

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:

130-112-791, 130-112-602

Protocols:

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

Description:

A mixture of human CD271 (LNGFR)–transfected Jurkat cells and human peripheral blood mononuclear cells (PBMCs) were stained with CD271 (LNGFR) antibodies or with the corresponding REA Control (S) antibodies (left peak) as well as with CD45 antibodies. CD45^– cells were pre-gated for the analysis. Flow cytometry was performed 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 forCD271 (LNGFR) Antibody, anti-human, REAfinity™

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-112-791, 130-112-602

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-112-791, 130-112-602

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD271 (LNGFR) 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 REA844
ME20.4-1.H4	++
ME20.4	++
C40-1457	+
REA648 (h/m)	-
REAL709	+
Cells were incubated with an excess of purified unconjugated CD271 (LNGFR) (REA844) 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 CD271 (LNGFR). CD271 (LNGFR) trasfected positive Jurkat cells and negative peripheral blood monoclear cells (PBMCs) were stained with CD271 (LNGFR) antibodies and with a suitable counterstaining. As a control, CD271 (LNGFR) 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 CD271 (LNGFR). CD271 (LNGFR) trasfected positive Jurkat cells and negative peripheral blood monoclear cells (PBMCs) were stained with CD271 (LNGFR) antibodies and with a suitable counterstaining. As a control, CD271 (LNGFR) 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 CD271 (LNGFR). CD271 (LNGFR) trasfected positive Jurkat cells and negative peripheral blood monoclear cells (PBMCs) were stained with CD271 (LNGFR) antibodies and with a suitable counterstaining. As a control, CD271 (LNGFR) 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 CD271 (LNGFR). CD271 (LNGFR) trasfected positive Jurkat cells and negative peripheral blood monoclear cells (PBMCs) were stained with CD271 (LNGFR) antibodies and with a suitable counterstaining. As a control, CD271 (LNGFR) 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 CD271 (LNGFR). CD271 (LNGFR) trasfected positive Jurkat cells and negative peripheral blood monoclear cells (PBMCs) were stained with CD271 (LNGFR) antibodies and with a suitable counterstaining. As a control, CD271 (LNGFR) 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 CD271 (LNGFR). CD271 (LNGFR) trasfected positive Jurkat cells and negative peripheral blood monoclear cells (PBMCs) were stained with CD271 (LNGFR) antibodies and with a suitable counterstaining. As a control, CD271 (LNGFR) 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 CD271 (LNGFR). CD271 (LNGFR) trasfected positive Jurkat cells and negative peripheral blood monoclear cells (PBMCs) were stained with CD271 (LNGFR) antibodies and with a suitable counterstaining. As a control, CD271 (LNGFR) 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 CD271 (LNGFR) (REA844). 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 CD271 (LNGFR) (REA844). 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 CD271 (LNGFR) Antibody, anti-human, REAfinity™

Overview

Clone REA844 recognizes the human CD271 antigen, also known as low-affinity nerve growth factor receptor (LNGFR) or p75 neurotrophin receptor (NTR). CD271 belongs to the low-affinity neurotrophin receptors and the tumor necrosis factor receptor superfamily. Initially the human CD271 (LNGFR) was described to be expressed on cells of the central and peripheral nervous system and was suggested to be involved in the development, survival, and differentiation of cells.

Expression of CD271 (LNGFR) is found on mesenchymal stem/stromal cells (MSCs), follicular dendritic cells, mesenchymal cells involved in mesenchymal-epithelial interactions, autonomic and sensory neurons, oligodendrocytes, astrocytes, schwann cells, and neural crest stem cells.

Additional information: Clone REA844 displays negligible binding to Fc receptors.

Alternative names

NGFR, Gp80-LNGFR, TNFRSF16, p75(NTR), p75NTR, p75NGFR, NGFR (p75)

Detailed product information

Technical specifications

Clone	REA844
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, non-human primate, other
Cross-reactivity	monkey, sheep, cat, dog, pig, rabbit
Antigen	CD271 (LNGFR)
Alternative names of antigen	NGFR, Gp80-LNGFR, TNFRSF16, p75(NTR), p75NTR, p75NGFR, NGFR (p75)
Molecular mass of antigen [kDa]	43
Distribution of antigen	stem cells, dendritic cells, epithelial cells, oligodendrocytes, astrocytes, Schwann cells, mesenchymal stem cells
Entrez Gene ID	4804
RRID	AB_2928367, AB_2928366, AB_2725868, AB_2725887, AB_2725864, AB_2725888, AB_2725865, AB_2725892, AB_2725869, AB_2725893, AB_2725870, AB_2725885, AB_2725862, AB_2725889, AB_2725866, AB_2725890, AB_2725867, AB_2725886, AB_2725863, AB_2725894, AB_2725871, AB_2725891

Resources for CD271 (LNGFR) Antibody, anti-human, REAfinity™

Documents and Protocols

Scientific posters

Sorting of MSCs with CD271 and MSCA-1 antibodies in a closed-cartridge system using the MACSQuant® Tyto® Sorter

Scientific posters

Standardized quantification of human bone marrow mesenchymal stromal cells based on a flow cytometry assay

Certificates

Please follow this

link

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

References for CD271 (LNGFR) Antibody, anti-human, REAfinity™

Publications

Caneva, L. et al. (1995) Immuno-electron microscopy characterization of human bone marrow stromal cells with anti-NGFR antibodies. Blood Cells Mol. Dis. 21: 73-85
Quirici, N. et al. (2002) Isolation of bone marrow mesenchymal stem cells by anti-nerve growth factor receptor antibodies. Exp. Hematol. 30: 783-791
Casaccia-Bonnefil, P. et al. (1996) Death of oligodendrocytes mediated by the interaction of nerve growth factor with its receptor p75. Nature 383: 716-719
DiStefano, P. S. and Johnson, E. M. Jr. (1988) Identification of a truncated form of the nerve growth factor receptor. Proc. Natl. Acad. Sci. U.S.A. 85: 270-274
Huber, L. J. and Chao, M. V. (1995) Mesenchymal and neuronal cell expression of the p75 neurotrophin receptor gene occur by different mechanisms. Dev. Biol. 167: 227-238
Chalazonitis, A. et al. (1998)
Age-dependent differences in the effects of GDNF and NT-3 on the development of neurons and glia from neural crest-derived precursors immunoselected from the fetal rat gut: expression of GFRalpha-1
in vitro
and
in vivo
.
Dev. Biol. 204(2): 385-406
Jones, E. A. et al. (2002) Isolation and characterization of bone marrow multipotential mesenchymal progenitor cells. Arthritis Rheum 46: 3349-3360
Kashiba, H. et al. (1995) Coexpression of trk family members and low-affinity neurotrophin receptors in rat dorsal root ganglion neurons. Brain Res. Mol. Brain Res. 30: 158-164
Rudge, J. S. et al. (1994) Neurotrophic factor receptors and their signal transduction capabilities in rat astrocytes. Eur. J. Neurosci. 6: 693-705
Thomson, T. M. et al. (1988) Expression of human nerve growth factor receptor on cells derived from all three germ layers. Exp. Cell Res. 174: 533-539
Pezzati, P. et al. (1992) Expression of nerve growth factor receptor immunoreactivity on follicular dendritic cells from human mucosa associated lymphoid tissues. Immunology 76: 485-490
Rozemuller, H. et al. (2010) Prospective isolation of mesenchymal stem cells from multiple mammalian species using cross-reacting anti-human monoclonal antibodies. Stem Cells Dev. 19: 1911-1921
Vroemen, M. and Weidner, N. (2003) Purification of Schwann cells by selection of p75 low affinity nerve growth factor receptor expressing cells from adult peripheral nerve. J. Neurosci. Methods 124: 135-143

Applications

Extended validation