CD45RO Antibody, anti-human,
REAlease
^®

Name: CD45RO Antibody, anti-human, REAlease®
Availability: InStock

CD45RO Antibody, anti-human,
REAlease
^®
Extended ValidationRecombinant

Clone:

REAL377

Type of antibody:

Releasable antibodies, Primary antibodies, Recombinant antibodies

Applications:

FC, MICS, IF, IHC

Flow cytometry applications forCD45RO Antibody, anti-human,
REAlease
^®

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Tested:

QC tested

Products used:

130-120-962, 130-120-874, 130-124-841

Protocols:

Cell surface staining protocol using REAlease™ antibodies

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with REAlease CD45RO Complexes or with the corresponding REAL Control (left images) as well as with CD45RA antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide (PI) fluorescence.

MICS (MACSima Imaging Cyclic Staining) applications forCD45RO Antibody, anti-human,
REAlease
^®

APC
PE

Conjugate:

APC

Recommended dilution:

1:12,5

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-120-962, 130-120-874

Tissue used:

spleen

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Description:

Immunofluorescence analysis of a human spleen cryosection fixed with ice-cold acetone. The section was stained using CD45RO Antibody, anti-human, APC (green) for 10 min at room temperature (A). CD45RO Antibody, anti-human, APC staining was released using the REAlease Release Reagent (B). Nuclei were visualized with Hoechst™ (blue). Scale bar: 30 µm.

Conjugate:

APC

Recommended dilution:

1:25

Fixation:

FFPE

Tested:

QC tested

Species tested:

human

Products used:

130-120-962, 130-120-874

Tissue used:

spleen

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Description:

Immunofluorescence analysis of a formalin-fixed, paraffin embedded (FFPE) human spleen. Before staining, antigens were retrieved using heat-induced epitope retrieval (HIER) at pH 9. The section was stained using CD45RO Antibody, anti-human, APC (green) for 10 min at room temperature (A). CD45RO Antibody, anti-human, APC staining was released using the REAlease Release Reagent (B). Nuclei were visualized with Hoechst™ (blue).Scale bar: 30 µm.

Extended validation for CD45RO Antibody, anti-human,
REAlease
^®

Specificity

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 CD45RO knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD45RO-PE (REAL377, red) and counterstained with DRAQ5 (blue) as DNA stain.

Fluorescence microscopy image of CD45RO knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD45RO-PE (REAL377, red) and counterstained with DRAQ5 (blue) as DNA stain.

Overlay histogram showing flow cytometric analysis of human CD45RO knockout cells. Wild type (red) and knockout cells (blue) were stained with CD45RO-PE, clone (REAL377). 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 CD45RO knockout cells. Wild type (red) and knockout cells (blue) were stained with CD45RO-PE, clone (REAL377). 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.

Specifications for CD45RO Antibody, anti-human,
REAlease
^®

Overview

Clone REAL377 is an antibody fragment derived from the full CD45RO antibody molecule. It displays no binding to Fc receptors. The recombinantly engineered antibody fragments are multimerized to form the REAlease Complex to bind markers with high avidity.

Clone REAL377 recognizes the human CD45RO antigen, a 180 kDa single-chain membrane glycoprotein, which is an isoform of CD45. CD45RO is expressed by most thymocytes, activated and memory T cells, granulocytes, B cell subsets, and monocytes. CD45RO binds to galectin-1 and CD22. CD45RO antibodies are commonly used in combination with antibodies against CD45RA to discriminate memory T cells from naive T cells.

The REAlease Kits consist of the respective fluorochrome-conjugated REAlease Complexes and the REAlease Support Kit for removal of the REAlease Complexes and optional relabeling with different fluorochrome-conjugated REAlease Complexes.

Detailed product information

Technical specifications

Clone	REAL377
Clonality	monoclonal
Isotype control	Control Antibody
Host	human cell line
Type of antibody	Releasable antibodies, Primary antibodies, Recombinant antibodies
Species	human
Antigen	CD45RO
Distribution of antigen	granulocytes, T cells, B cells, monocytes, thymocytes
RRID	AB_2811683, AB_2784223, AB_2784222, AB_2784221, AB_2811684, AB_2784224

Resources for CD45RO Antibody, anti-human,
REAlease
^®

Certificates

Please follow this

link

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

Applications

Extended validation