CD90 Antibody, anti-human, REAfinity™

Name: CD90 Antibody, anti-human, REAfinity™
Availability: InStock

CD90 Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA897

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC

Alternative names:

Thy-1, THY1, CDw90

Flow cytometry applications forCD90 Antibody, anti-human, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700
Vio Bright B515
Vio Bright R720
VioBlue

Conjugate:

APC

Recommended dilution:

1:50

Remark:

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

Tested:

QC tested

Products used:

130-114-903, 130-114-861

Protocols:

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

Description:

HEL 92.1.7 cells were stained with CD90 antibodies or with the corresponding REA Control (S) antibodies (left peak). 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 forCD90 Antibody, anti-human, REAfinity™

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-114-903, 130-114-861

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD90 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 REA897
5E10	++
F15-42-1	++
DG3	+
REAL449	+
Cells were incubated with an excess of purified unconjugated CD90 (REA897) 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.

Knockout validation

To ensure antibody specificity, the target gene is knocked out in a suitable cell line using the CRISPR/Cas9 system and the knockout is confirmed by sequencing of the target locus. The antibody is considered to bind specifically to the intended epitope if no antibody binding to the knockout cells can be detected. The antibody staining is controlled by fluorescence microscopy and/or flow cytometry.

WT

KO

Fluorescence microscopy image of CD90 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD90-PE (REA897, red) and counterstained with DRAQ5 (blue) as DNA stain.

Fluorescence microscopy image of CD90 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD90-PE (REA897, red) and counterstained with DRAQ5 (blue) as DNA stain.

Overlay histogram showing flow cytometric analysis of CD90 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD90-PE, clone (REA897). 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 propidium iodide fluorescence.

Overlay histogram showing flow cytometric analysis of CD90 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD90-PE, clone (REA897). 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 propidium iodide fluorescence.

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 CD90. HEL 92.1.7 cells were stained with CD90 antibodies and with a suitable counterstaining. As a control, CD90 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 CD90. HEL 92.1.7 cells were stained with CD90 antibodies and with a suitable counterstaining. As a control, CD90 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 CD90. HEL 92.1.7 cells were stained with CD90 antibodies and with a suitable counterstaining. As a control, CD90 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 CD90. HEL 92.1.7 cells were stained with CD90 antibodies and with a suitable counterstaining. As a control, CD90 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 CD90. HEL 92.1.7 cells were stained with CD90 antibodies and with a suitable counterstaining. As a control, CD90 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 CD90. HEL 92.1.7 cells were stained with CD90 antibodies and with a suitable counterstaining. As a control, CD90 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 CD90. HEL 92.1.7 cells were stained with CD90 antibodies and with a suitable counterstaining. As a control, CD90 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 CD90. HEL 92.1.7 cells were stained with CD90 antibodies and with a suitable counterstaining. As a control, CD90 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 CD90. HEL 92.1.7 cells were stained with CD90 antibodies and with a suitable counterstaining. As a control, CD90 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 CD90 (REA897). 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 CD90 (REA897). 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 CD90 (REA897). 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 CD90 (REA897). 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 CD90 Antibody, anti-human, REAfinity™

Overview

Clone REA897 recognizes the human CD90 antigen, which is a 25–35 kD GPI-anchored protein of the Ig superfamily. It is expressed on neurons, small subsets of human fetal liver cells and thymocytes, cord blood, and bone marrow cells. CD90 is also expressed on a subset of CD34

⁺

primitive hematopoietic stem cells. CD90

⁺

CD34

⁺

cells characterize a highly enriched population of high proliferative potential colony-forming cells (HPP-CFC). Furthermore, CD90 expression is found on mesenchymal stromal cells (MSCs). CD90, also known as Thy-1, is involved in regulation of adhesion and signal transduction of T cells.

Additional information: Clone REA897 displays negligible binding to Fc receptors.

Alternative names

Thy-1, THY1, CDw90

Detailed product information

Technical specifications

Clone	REA897
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
Cross-reactivity	cynomolgus monkey ( Macaca fascicularis ) , rhesus monkey ( Macaca mulatta ) , baboon, pigtailed macaque
Antigen	CD90
Alternative names of antigen	Thy-1, THY1, CDw90
Molecular mass of antigen [kDa]	13
Distribution of antigen	stem cells, endothelial cells, fibroblasts, leukocytes, lymphocytes, neurons, ILC, T cells, cancer stem cells, hematopoietic stem and progenitor cells, leukemia cells, mesenchymal stem cells, ES and iPS cells, thymocytes, brain
Entrez Gene ID	7070
RRID	AB_2726810, AB_2726842, AB_2726811, AB_2726843, AB_2726812, AB_2726847, AB_2726816, AB_2726848, AB_2726817, AB_2726844, AB_2726813, AB_2726845, AB_2726814, AB_2726846, AB_2726815, AB_2905143, AB_2905142, AB_2726809, AB_2928581, AB_2928580, AB_2726841

Resources for CD90 Antibody, anti-human, REAfinity™

Certificates

Please follow this

link

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

References for CD90 Antibody, anti-human, REAfinity™

Publications

Mayani, H. and Lansdorp, P. M. (1994) Thy-1 expression is linked to functional properties of primitive hematopoietic progenitor cells from human umbilical cord blood. Blood 83: 2410-2417
Wetzel, A. et al. (2004) Human Thy-1 (CD90) on activated endothelial cells is a counterreceptor for the leukocyte integrin Mac-1 (CD11b/CD18). J Immunol 172(6): 3850-3859
Dominici, M. et al. (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8: 315-317

Applications

Extended validation