Clone:
REA747
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, ICC, MC, 3D-IF
Alternative names:
Ptprc, GP180, L-CA, LY5, T200

Extended validation for CD45 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 REA747
5B1++
2D1++
HI30++
MB4-6D6-
REA1023-
REAL258++
Cells were incubated with an excess of purified unconjugated CD45 (REA747) 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 CD45 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD45-PE (REA747, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Fluorescence microscopy image of CD45 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD45-PE (REA747, red) and counterstained with DRAQ5 (blue) as DNA stain.
Fluorescence microscopy image of CD45 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD45-PE (REA747, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Overlay histogram showing flow cytometric analysis of human CD45 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD45-PE, clone (REA747). 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 CD45 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD45-PE, clone (REA747). 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 CD45. Human peripheral blood mononuclear cells (PBMCs) were stained with CD45 antibodies and plotted against the side scatter. As a control, CD45 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 propidium iodide 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 CD45. Human peripheral blood mononuclear cells (PBMCs) were stained with CD45 antibodies and plotted against the side scatter. As a control, CD45 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 propidium iodide 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 CD45. Human peripheral blood mononuclear cells (PBMCs) were stained with CD45 antibodies and plotted against the side scatter. As a control, CD45 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 propidium iodide 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 CD45. Human peripheral blood mononuclear cells (PBMCs) were stained with CD45 antibodies and plotted against the side scatter. As a control, CD45 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 propidium iodide 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 CD45. Human peripheral blood mononuclear cells (PBMCs) were stained with CD45 antibodies and plotted against the side scatter. As a control, CD45 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 propidium iodide 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 CD45. Human peripheral blood mononuclear cells (PBMCs) were stained with CD45 antibodies and plotted against the side scatter. As a control, CD45 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 propidium iodide 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 CD45. Human peripheral blood mononuclear cells (PBMCs) were stained with CD45 antibodies and plotted against the side scatter. As a control, CD45 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 propidium iodide fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for CD45 Antibody, anti-human, REAfinity™

Overview

Clone REA747 recognizes the human CD45 antigen, a member of the protein tyrosine phosphatase (PTP) family, also known as the leukocyte common antigen (LCA). The CD45 molecule is a type I transmembrane protein which is required for T and B cell activation and involved in cell growth, differentiation, mitotic cycle, and oncogenic transformation. CD45 is expressed in different isoforms (CD45RA, CD45RB, CD45RC, CD45RAB, CD45RAC, CD45RBC, CD45RO, CD45R (ABC)) depending on the differentiation status of the cell. The CD45 antibody recognizes a common epitope of all CD45 isoforms.
Additional information: Clone REA747 displays negligible binding to Fc receptors.

Alternative names

Ptprc, GP180, L-CA, LY5, T200

Detailed product information

Technical specifications

CloneREA747
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD45
Alternative names of antigenPtprc, GP180, L-CA, LY5, T200
Molecular mass of antigen [kDa]145
Distribution of antigenT cells, B cells, dendritic cells, Langerhans cells, mast cells, monocytes, macrophages, leukocytes, lymphocytes, thymocytes, hematopoietic stem and progenitor cells
Entrez Gene ID5788
RRIDAB_2658237, AB_2726700, AB_2658238, AB_2658239, AB_2658240, AB_2658241, AB_2658242, AB_2658243, AB_2658244, AB_2658245, AB_2658246, AB_2658247, AB_2658248, AB_2658249, AB_2658250, AB_2658251, AB_2658252, AB_2658253, AB_2658254, AB_2658255, AB_2751047, AB_2751046, AB_2658256, AB_2658257, AB_2801892, AB_2658236

Resources for CD45 Antibody, anti-human, REAfinity™

Certificates

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

References for CD45 Antibody, anti-human, REAfinity™

Publications

  1. Ralph, S. J. et al. (1987) Structural variants of human T200 glycoprotein (leukocyte-common antigen). EMBO J. 6(5): 1251-1257
  2. Vykoukal, J. et al. (2008) Enrichment of putative stem cells from adipose tissue using dielectrophoretic field-flow fractionation. Lab Chip 8(8): 1386-1393
  3. Kurian, L. et al. (2013) Conversion of human fibroblasts to angioblast-like progenitor cells. Nat. Methods 10(1): 77-83

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