CD45RA Antibody, anti-human,
REAlease
^®

Name: CD45RA Antibody, anti-human, REAlease®
Availability: InStock

CD45RA Antibody, anti-human,
REAlease
^®
Extended ValidationRecombinant

Clone:

REAL164

Type of antibody:

Releasable antibodies, Primary antibodies, Recombinant antibodies

Applications:

FC, MICS, IHC, IF

Alternative names:

Ptprc, B220, CD45, CD45R, GP180, L-CA, LY5, T200

Flow cytometry applications forCD45RA Antibody, anti-human,
REAlease
^®

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Tested:

QC tested

Products used:

130-112-264, 130-112-097

Protocols:

Cell surface staining protocol using REAlease™ antibodies

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with REAlease CD45RA Complexes or with the corresponding REAL Control (left images) as well as with CD45RO 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 forCD45RA Antibody, anti-human,
REAlease
^®

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

FFPE

Tested:

during development

Species tested:

human

Products used:

130-112-264, 130-112-097

Tissue used:

tonsil

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Description:

Immunofluorescence analysis of a formalin-fixed, paraffin embedded (FFPE) human tonsil. Before staining, antigens were retrieved using heat-induced epitope retrieval (HIER) at pH 9. The section was stained using CD45RA Antibody, anti-human, APC (green) for 10 min at room temperature (A). CD45RA 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:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-112-264, 130-112-097

Tissue used:

tonsil

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Description:

Immunofluorescence analysis of a human tonsil cryosection fixed with 4% PFA. The section was stained using CD45RA Antibody, anti-human, APC (green) for 10 min at room temperature (A). CD45RA 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:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-112-264, 130-112-097

Tissue used:

tonsil

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD45RA 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 CD45RA knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD45RA-PE (REAL164, red) and counterstained with DRAQ5 (blue) as DNA stain.

Fluorescence microscopy image of CD45RA knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD45RA-PE (REAL164, red) and counterstained with DRAQ5 (blue) as DNA stain.

Overlay histogram showing flow cytometric analysis of CD45RA knockout cells. Wild type (red) and knockout cells (blue) were stained with CD45RA-PE, clone (REA164). 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 CD45RA knockout cells. Wild type (red) and knockout cells (blue) were stained with CD45RA-PE, clone (REA164). 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 CD45RA Antibody, anti-human,
REAlease
^®

Overview

Clone REAL164 is an antibody fragment derived from the full CD45RA 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 REAL164 recognizes the human CD45RA antigen which is expressed on naive CD4

⁺

and CD8

⁺

T cells as well as on CD8

⁺

effector T cells. CD45RA is also present on subsets of B and NK cells and on plasmacytoid dendritic cells. The CD45RA antibody recognizes the 220 kDa isoform of the leukocyte common antigen (LCA), a transmembrane tyrosine phosphatase.

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.

Alternative names

Ptprc, B220, CD45, CD45R, GP180, L-CA, LY5, T200

Detailed product information

Technical specifications

Clone	REAL164
Clonality	monoclonal
Isotype control	Control Antibody
Host	cell line
Type of antibody	Releasable antibodies, Primary antibodies, Recombinant antibodies
Species	human
Cross-reactivity	cynomolgus monkey ( Macaca fascicularis ) , rhesus monkey ( Macaca mulatta )
Antigen	CD45RA
Alternative names of antigen	Ptprc, B220, CD45, CD45R, GP180, L-CA, LY5, T200
Distribution of antigen	T cells, B cells, NK cells, dendritic cells, CD4+ T cells, CD8+ T cells, plasmacytoid dendritic cells
RRID	AB_2751966, AB_2751920, AB_2819366, AB_2819365, AB_2784220, AB_2751072

Resources for CD45RA Antibody, anti-human,
REAlease
^®

Certificates

Please follow this

link

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

References for CD45RA Antibody, anti-human,
REAlease
^®

Publications

Cantisan, S. et al. (2010)
CD45RA expression on HCMV-specific effector memory CD8
⁺
T cells is associated with the duration and intensity of HCMV replication after transplantation.
Clin. Immunol. 137(1): 81-88
Baba, N. et al. (2010)
Interplay between CD45RA
⁺
regulatory T cells and TNF-α in the regulation of human Tʜ17 differentiation.
Int. Immunol. 22: 237-244
Braun, T. et al. (2012)
Effector CD4
⁺
CD45RA
^–
CD25
^bright
Foxp3
^bright
regulatory T cell (eTreg) distribution is significantly impaired in chronic myelomonocytic leukemia (CMML) and correlates with TET 2 mutational status.
Blood 120: 2808-2808

Applications

Extended validation