Basophils Antibody, anti-human, REAfinity™

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

Basophils Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA567

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

ICFC

Intracellular flow cytometry applications forBasophils Antibody, anti-human, REAfinity™

APC
PE
Vio Bright B515

Conjugate:

APC

Recommended dilution:

1:11

Remark:

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

Tested:

QC tested

Products used:

130-108-845, 130-108-819

Protocols:

Intracellular flow cytometry staining protocol (Cell Signaling Buffer Set A 1:11)

Description:

Human peripheral blood mononuclear cells (PBMCs) were fixed and permeabilized using the Cell Signaling Buffer Set A. Cells were then stained with Anti-Basophils antibodies or with the corresponding REA control (I) antibodies (left image) as well as with CD193 antibodies. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris were excluded from the analysis based on scatter signals.

Extended validation for Basophils 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 REA567
2D7/Baso	++
Bsp-1	-
Basophil/2D7	++
2D7	++
Cells were incubated with an excess of purified unconjugated Basophils (REA567) 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 Basophils. Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Cell Signaling Buffer Set A followed by intracellular staining with Basophils antibodies. As a control, Basophils 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 Viobility 405/520 Fixable Dye 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 Basophils. Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Cell Signaling Buffer Set A followed by intracellular staining with Basophils antibodies. As a control, Basophils 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 Viobility 405/520 Fixable Dye 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 Basophils. Human peripheral blood mononuclear cells (PBMCs) were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Cell Signaling Buffer Set A followed by intracellular staining with Basophils antibodies. As a control, Basophils 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 Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for Basophils Antibody, anti-human, REAfinity™

Overview

Clone REA567 recognizes the population of human basophils. Basophils appear in many specific kinds of inflammatory reactions, particularly those that cause allergic symptoms, like parasitic infections and allergies. They secrete a wide variety of proinflammatory agents upon activation such as histamine and cytokines.

Additional information: Clone REA567 displays negligible binding to Fc receptors.

Detailed product information

Technical specifications

Clone	REA567
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (I), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human
Antigen	Basophils
Distribution of antigen	basophils
RRID	AB_2819432, AB_2651254, AB_2651255, AB_2651256, AB_2651257, AB_2819433

Resources for Basophils Antibody, anti-human, REAfinity™

Certificates

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References for Basophils Antibody, anti-human, REAfinity™

Publications

Schroeder, J. T. (2009) Basophils beyond effector cells of allergic inflammation. Adv. Immunol. 101: 123-161
Nakanishi, K. (2010) Basophils as APC in Th2 response in allergic inflammation and parasite infection. Curr. Opin. Immunol. 22(6): 814-820

Applications

Extended validation