Clone:
REA263
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, IF, IHC, MICS, MC
Alternative names:
ANPEP, APN, gp150, LAP1, p150, PEPN

Extended validation for CD13 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 REA263
L138-
WM15++
TUK1-
REAL312++
Cells were incubated with an excess of purified unconjugated CD13 (REA263) 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 CD13 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD13-PE (REA263, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Fluorescence microscopy image of CD13 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD13-PE (REA263, red) and counterstained with DRAQ5 (blue) as DNA stain.
Fluorescence microscopy image of CD13 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD13-PE (REA263, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Overlay histogram showing flow cytometric analysis of CD13 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD13-PE, clone (REA263). 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 CD13 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD13-PE, clone (REA263). 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 CD13. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD13 antibodies and plotted against the side scatter. As a control, CD13 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 CD13. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD13 antibodies and plotted against the side scatter. As a control, CD13 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 CD13. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD13 antibodies and plotted against the side scatter. As a control, CD13 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 CD13. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD13 antibodies and plotted against the side scatter. As a control, CD13 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 CD13. Human peripheral blood mononuclear cells (PBMCs) after erythrocyte lysis were stained with CD13 antibodies and plotted against the side scatter. As a control, CD13 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 CD13 (REA263). 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 CD13 (REA263). 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 CD13 (REA263). 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 CD13 Antibody, anti-human, REAfinity™

Overview

Clone REA263 recognizes the CD13 antigen, a 150–170 kDa type II transmembrane glycoprotein, which is also known as aminopeptidase N or gp150. CD13 is expressed on granulocytes, myeloid progenitors, endothelial cells, epithelial cells, and subset of granular lymphoid cells. It is also broadly expressed in other tissues such as kidney proximal tubules, intestine, and placenta. CD13 is an enzyme that is used as a biomarker to detect damage to the kidneys, and that may be used to help diagnose certain kidney disorders. It also serves as a receptor for one strain of human coronavirus that is an important cause of upper respiratory tract infections. Defects in CD13 appear to be a cause of various types of leukemia or lymphoma.
Additional information: Clone REA263 displays negligible binding to Fc receptors.

Alternative names

ANPEP, APN, gp150, LAP1, p150, PEPN

Detailed product information

Technical specifications

CloneREA263
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD13
Alternative names of antigenANPEP, APN, gp150, LAP1, p150, PEPN
Molecular mass of antigen [kDa]109
Distribution of antigenendothelial cells, granulocytes, epithelial cells
Entrez Gene ID290
RRIDAB_2783936, AB_2751875, AB_2751845, AB_2751792, AB_2751732, AB_2819547, AB_2819512, AB_2811584, AB_2811560, AB_2654853, AB_2654854, AB_2783934, AB_2783937, AB_2783935

Resources for CD13 Antibody, anti-human, REAfinity™

Certificates

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

References for CD13 Antibody, anti-human, REAfinity™

Publications

  1. Look, A. T. et al. (1989) Human myeloid plasma membrane glycoprotein CD13 (gp150) is identical to aminopeptidase N. J. Clin. Invest. 83(4): 1299-1307
  2. Favaloro, E. J. et al. (1993) CD13 (GP150; aminopeptidase-N): predominant functional activity in blood is localized to plasma and is not cell-surface associated. Exp. Hematol. 21(13): 1695-1701
  3. Yeager, C. L. et al. (1992) Human aminopeptidase N is a receptor for human coronavirus 229E. Nature 357(6377): 420-422

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