CD102 (ICAM-2) Antibody, anti-human, REAfinity™

Name: CD102 (ICAM-2) Antibody, anti-human, REAfinity™
Availability: InStock

CD102 (ICAM-2) Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA878

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC

Alternative names:

ICAM-2

Flow cytometry applications forCD102 (ICAM-2) Antibody, anti-human, REAfinity™

APC
APC-Vio 770
PE
PE-Vio 770
Vio Bright V423

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:

130-114-412, 130-114-305

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD102 (ICAM-2) antibodies or with the corresponding REA Control (S) antibodies (left image) as well as with CD45 antibodies. 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.

Extended validation for CD102 (ICAM-2) 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 REA878
CBR-IC2/2	++
Cells were incubated with an excess of purified unconjugated CD102 (ICAM-2) (REA878) 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 CD102 (ICAM-2) knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD102 (ICAM-2)-PE (REA878, red) and counterstained with DRAQ5 (blue) as DNA stain.

Fluorescence microscopy image of CD102 (ICAM-2) knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD102 (ICAM-2)-PE (REA878, red) and counterstained with DRAQ5 (blue) as DNA stain.

Overlay histogram showing flow cytometric analysis of human CD45RB knockout cells. Wild type (red) and knockout cells (blue) were stained with CD45RB-PE, clone (REA119). 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 human CD45RB knockout cells. Wild type (red) and knockout cells (blue) were stained with CD45RB-PE, clone (REA119). 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 CD102 (ICAM-2). Human peripheral blood mononuclear cells (PBMCs) were stained with CD102 (ICAM-2) antibodies and with a suitable counterstaining. As a control, CD102 (ICAM-2) 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 CD102 (ICAM-2). Human peripheral blood mononuclear cells (PBMCs) were stained with CD102 (ICAM-2) antibodies and with a suitable counterstaining. As a control, CD102 (ICAM-2) 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 CD102 (ICAM-2). Human peripheral blood mononuclear cells (PBMCs) were stained with CD102 (ICAM-2) antibodies and with a suitable counterstaining. As a control, CD102 (ICAM-2) 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 CD102 (ICAM-2) (REA878). 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 CD102 (ICAM-2) (REA878). 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 CD102 (ICAM-2) Antibody, anti-human, REAfinity™

Overview

Clone REA878 recognizes the human CD102 antigen, a type I transmembrane glycoprotein with two immunoglobulin-like domains. CD102, also known as ICAM-2, is a member of the intercellular adhesion molecule (ICAM) family, that binds to leukocyte integrin, lymphocyte function-associated antigen-1 (LFA-1), dendritic cell-specific antigen, ICAM-grabbing non-integrin (DC-SIGN) protein and macrophage-1 antigen (MAC-1). CD102 is found on vascular endothelial cells, monocytes, platelets and different populations of lymphocytes..

CD102 is involved in lymphocyte recirculation and induces essential adhesive cell interactions, which are important for immune response and surveillance.

Additional information: Clone REA878 displays negligible binding to Fc receptors.

Alternative names

ICAM-2

Detailed product information

Technical specifications

Clone	REA878
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
Antigen	CD102 (ICAM-2)
Alternative names of antigen	ICAM-2
Molecular mass of antigen [kDa]	55
Distribution of antigen	endothelial cells, monocytes, platelets, lymphocytes
Entrez Gene ID	3384
RRID	AB_2726616, AB_2726547, AB_2726617, AB_2726548, AB_2726618, AB_2726549, AB_2726619, AB_2726550, AB_2921914, AB_2921883

Resources for CD102 (ICAM-2) Antibody, anti-human, REAfinity™

Certificates

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References for CD102 (ICAM-2) Antibody, anti-human, REAfinity™

Publications

Staunton, D. E. et al. (1989) Functional cloning of ICAM-2, a cell adhesion ligand for LFA-1 homologous to ICAM-1. Nature 339(6219): 61-64
Simmons, D. L. (1995) The role of ICAM expression in immunity and disease. Cancer Surv. 24: 144-155
Weber, K. S. et al. (2004) Sialylation of ICAM-2 on platelets impairs adhesion of leukocytes via LFA-1 and DC-SIGN. Inflammation 28(4): 177-188

Applications

Extended validation