CD41a Antibody, anti-human, REAfinity™

Name: CD41a Antibody, anti-human, REAfinity™
Availability: InStock

CD41a Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA386

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC

Alternative names:

ITGA2B, BDPLT16, BDPLT2, GP2B, GPIIb

Flow cytometry applications forCD41a Antibody, anti-human, REAfinity™

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

Conjugate:

APC

Recommended dilution:

1:50

Remark:

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

Tested:

QC tested

Products used:

130-128-897, 130-128-830

Protocols:

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

Description:

Human peripheral blood platelets were stained with CD41a antibodies or with the corresponding REA Control (S) antibodies (left peak) and analyzed by flow cytometry using the MACSQuant^® Analyzer. Platelets were pre-gated for the analysis based on forward and side light-scatter characteristics. 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 forCD41a Antibody, anti-human, REAfinity™

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-128-897, 130-128-830

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD41a 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 REA386
HIP8	++
CLB-tromb, 6C9	+
Cells were incubated with an excess of purified unconjugated CD41a (REA386) 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 CD41a. Human peripheral blood platelets were stained with CD41a antibodies and with a suitable counterstaining. As a control, CD41a 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 CD41a. Human peripheral blood platelets were stained with CD41a antibodies and with a suitable counterstaining. As a control, CD41a 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 CD41a. Human peripheral blood platelets were stained with CD41a antibodies and with a suitable counterstaining. As a control, CD41a 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 CD41a (REA386). 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 CD41a (REA386). 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 CD41a Antibody, anti-human, REAfinity™

Overview

Clone REA386 recognizes the human CD41a antigen, a calcium-dependent complex of CD41/CD61 (GPIIb/IIIa) expressed on normal platelets and megakaryocytes. The CD41/CD61 complex is a receptor for fibronectin, fibrinogen, von Willebrand factor, vitronectin, and thrombospondin and is required for platelet aggregation and clotting. Mutations of either CD41 or CD61 in humans causes Glanzmann’s thrombasthenia. Notably, CD41 is also the first blood-specific marker during

in vitro

differentiation of embryonic stem cells and is expressed on all hematopoietic stem and progenitor cells in the early embryo. However, CD41 expression is downregulated in midgestation embryos, and it is currently thought that CD41 expression on adult hematopoietic stem cells remains switched off throughout adult life.

Additional information: Clone REA386 displays negligible binding to Fc receptors.

Alternative names

ITGA2B, BDPLT16, BDPLT2, GP2B, GPIIb

Detailed product information

Technical specifications

Clone	REA386
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	chimpanzee ( Pan troglodytes ) , baboon, capuchin monkey
Antigen	CD41a
Alternative names of antigen	ITGA2B, BDPLT16, BDPLT2, GP2B, GPIIb
Molecular mass of antigen [kDa]	110
Distribution of antigen	platelets, megakaryocytes, stem cells, hematopoietic stem and progenitor cells
Entrez Gene ID	3674
RRID	AB_2752178, AB_2784190, AB_2784189, AB_2752223, AB_2752222, AB_2819562, AB_2819532, AB_2857722, AB_2857717, AB_2658046, AB_2658047, AB_2658054, AB_2658055, AB_2921967, AB_2921950, AB_2905004, AB_2905003, AB_2922249, AB_2922213, AB_2752197

Resources for CD41a Antibody, anti-human, REAfinity™

Certificates

Please follow this

link

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

References for CD41a Antibody, anti-human, REAfinity™

Publications

Gekas, C. et al. (2013) CD41 expression marks myeloid-biased adult hematopoietic stem cells and increases with age. Blood 121(22): 4463-4472
Poncz, M. et al. (1987) Structure of the platelet membrane glycoprotein IIb. Homology to the alpha subunits of the vitronectin and fibronectin membrane receptors. J. Biol. Chem. 262(18): 8476-8482

Applications

Extended validation