Clone:
REA878
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
ICAM-2

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 clonesOverlap 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
View details
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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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
View details
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.
View details
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

CloneREA878
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD102 (ICAM-2)
Alternative names of antigenICAM-2
Molecular mass of antigen [kDa]55
Distribution of antigenendothelial cells, monocytes, platelets, lymphocytes
Entrez Gene ID3384
RRIDAB_2726616, AB_2726547, AB_2726617, AB_2726548, AB_2726618, AB_2726549, AB_2726619, AB_2726550

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

Publications

  1. 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
  2. Simmons, D. L. (1995) The role of ICAM expression in immunity and disease. Cancer Surv. 24: 144-155
  3. 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

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