Clone:
REA259
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, ICC
Alternative names:
MOX44, TSPAN25

Extended validation for CD53 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 REA259
HI29++
MEM-53+
Cells were incubated with an excess of purified unconjugated CD53 (REA259) 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 CD53 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD53-PE (REA259, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Fluorescence microscopy image of CD53 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD53-PE (REA259, red) and counterstained with DRAQ5 (blue) as DNA stain.
Fluorescence microscopy image of CD53 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD53-PE (REA259, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Overlay histogram showing flow cytometric analysis of CD53 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD53-PE, clone (REA259). 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 CD53 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD53-PE, clone (REA259). 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 CD53 Antibody, anti-human, REAfinity™

Overview

Clone REA259 recognizes the CD53 antigen, a 35–42 kDa type II transmembrane glycoprotein, which is also known as tetraspanin-25. CD53 is expressed on all leukocytes including plasma cells, but not on platelets, erythrocytes, and non-hematopoietic tissues. CD53 mediates oxidative burst in monocytes and has been shown to interact with a number of proteins including IL-4, CD20, CD2, CD9, IL-2, VLA-4, and CD4. Cross-linking of CD53 on monocytes induces a strong increase in intracellular calcium ions, whereas a modest increase is seen in B lymphocytes. In resting B cells, the molecule transmits a co-stimulatory signal and promotes activation through the Ig receptor.
Additional information: Clone REA259 displays negligible binding to Fc receptors.

Alternative names

MOX44, TSPAN25

Detailed product information

Technical specifications

CloneREA259
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD53
Alternative names of antigenMOX44, TSPAN25
Distribution of antigenB cells, monocytes, T cells
Entrez Gene ID963
RRIDAB_2658657, AB_2658658, AB_2658659, AB_2658660, AB_2658661, AB_2658662, AB_2658663, AB_2658664, AB_2658665, AB_2658666, AB_2658667, AB_2658656

Resources for CD53 Antibody, anti-human, REAfinity™

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

References for CD53 Antibody, anti-human, REAfinity™

Publications

  1. Olweus, J. et al. (1993) CD53, a protein with four membrane-spanning domains, mediates signal transduction in human monocytes and B cells. J Immunol 151: 707-716
  2. Maecker, H. T, et al. (1997) The tetraspanin superfamily: molecular facilitators. FASEB J. 11: 428-442
  3. Angelisova, P. et al. (1990) The human leucocyte surface antigen CD53 is a protein structurally similar to the CD37 and MRC OX-44 antigens. Immunogenetics 32(4): 281-285

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