CD282 (TLR2) Antibody, anti-human/mouse, REAfinity™

Name: CD282 (TLR2) Antibody, anti-human/mouse, REAfinity™
Availability: InStock

CD282 (TLR2) Antibody, anti-human/mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA109

Type of antibody:

Primary antibodies, Recombinant antibodies, Functional-grade antibodies

Isotype:

recombinant human IgG1

Applications:

FC, FA, MC

Alternative names:

TLR2, TIL4, Ly105

Flow cytometry applications forCD282 (TLR2) Antibody, anti-human/mouse, REAfinity™

APC
APC-Vio 770
Biotin
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700

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:

130-120-138, 130-120-052

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD282 (TLR2) antibodies as well as with CD14 antibodies and 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.

Functional assay applications forCD282 (TLR2) Antibody, anti-human/mouse, REAfinity™

pure-functional grade

Conjugate:

pure-functional grade

Products used:

130-098-855

Type of functional assay:

blocking of receptor-ligand binding

Mass cytometry applications forCD282 (TLR2) Antibody, anti-human/mouse, REAfinity™

pure

Conjugate:

pure

Products used:

130-108-058

Extended validation for CD282 (TLR2) Antibody, anti-human/mouse, 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 REA109
11G7	-
T2.5	++
TL2.1	+
QA16A01	+
W15145C	-
Cells were incubated with an excess of purified unconjugated CD282 (TLR2) (REA109) 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 CD282 (TLR2) knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD282 (TLR2)-PE (REA109, red) and counterstained with DRAQ5 (blue) as DNA stain.

Fluorescence microscopy image of CD282 (TLR2) knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD282 (TLR2)-PE (REA109, red) and counterstained with DRAQ5 (blue) as DNA stain.

Overlay histogram showing flow cytometric analysis of CD282 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD282-PE, clone (REA109). 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 CD282 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD282-PE, clone (REA109). 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 CD282 (TLR2). Human peripheral blood mononuclear cells (PBMCs) were stained with CD282 (TLR2) antibodies and with a suitable counterstaining. As a control, CD282 (TLR2) 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 CD282 (TLR2). Human peripheral blood mononuclear cells (PBMCs) were stained with CD282 (TLR2) antibodies and with a suitable counterstaining. As a control, CD282 (TLR2) 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 CD282 (TLR2). Human peripheral blood mononuclear cells (PBMCs) were stained with CD282 (TLR2) antibodies and with a suitable counterstaining. As a control, CD282 (TLR2) 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 CD282 (TLR2) (REA109). 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 CD282 (TLR2) (REA109). 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 CD282 (TLR2) Antibody, anti-human/mouse, REAfinity™

Overview

Clone REA109 recognises human and mouse CD282, also known as toll-like receptor 2 (TLR2). TLRs are the host pattern recognition receptors involved in recognising pathogen associated and certain tissue damage indicating molecules. They are type 1 transmembrane proteins with extracellular leucine-rich repeats and intracellular toll/IL-1 receptor domains. TLR2 is a plasma membrane associated TLR and recognizes various structurally diverse molecules including peptidoglycans (PG), lipopeptides and lipoproteins moieties in Gram

⁺

and Gram

^–

bacteria, lipoteichoic acid (LTA) in Gram

⁺

bacteria, lipoarabinomannan and lipomannan from Mycoplasma, and fungal polysaccharides such as zymogen. High-resolution crystal structures suggested that TLR2 by forming heterodimers with TLR6 and TLR1 can distinguish between di-acylated and tri-acylated lipopeptides. The expression of CD282 (TLR2) is found on monocytes, granulocytes, dendritic, endothelial, and epithelial cells.

Additional information: Clone REA109 displays negligible binding to Fc receptors.

Alternative names

TLR2, TIL4, Ly105

Detailed product information

Technical specifications

Clone	REA109
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, Functional-grade antibodies
Species	human, mouse
Antigen	CD282 (TLR2)
Alternative names of antigen	TLR2, TIL4, Ly105
Molecular mass of antigen [kDa]	88
Entrez Gene ID	7097
RRID	AB_2751976, AB_2889534, AB_2889533, AB_2661336, AB_2656979, AB_2656973, AB_2656974, AB_2656977, AB_2656978, AB_2904905, AB_2904904, AB_2904907, AB_2904906, AB_2921959, AB_2921939, AB_2752006

Resources for CD282 (TLR2) Antibody, anti-human/mouse, REAfinity™

Certificates

Please follow this

link

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

References for CD282 (TLR2) Antibody, anti-human/mouse, REAfinity™

Publications

Kang, J. Y. et al. (2009) Recognition of lipopeptide patterns by Toll-like receptor 2-Toll-like receptor 6 heterodimer. Immunity 31(6): 873-884
Takeda, K. and Akira, S. (2005) Toll-like receptors in innate immunity. Int. Immunol. 17: 1-14
Kirschning, C. J. and Schumann, R. R. (2002) TLR2: cellular sensor for microbial and endogenous molecular patterns. Curr Top Microbiol Immunol. 270: 121-144

Applications

Extended validation