Clone:
REA164
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
CDW52, CAMPATH-1

Extended validation for CD52 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 REA164
HI186n/a
4C8+
Hu116 (Campath-1H)++
QA19A22++
YTH34.5++
Cells were incubated with an excess of purified unconjugated CD52 (REA164) 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.

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 CD52. Human peripheral blood mononuclear cells (PBMCs) were stained with CD52 antibodies and with a suitable counterstaining. As a control, CD52 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 CD52. Human peripheral blood mononuclear cells (PBMCs) were stained with CD52 antibodies and with a suitable counterstaining. As a control, CD52 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 CD52. Human peripheral blood mononuclear cells (PBMCs) were stained with CD52 antibodies and with a suitable counterstaining. As a control, CD52 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 CD52. Human peripheral blood mononuclear cells (PBMCs) were stained with CD52 antibodies and with a suitable counterstaining. As a control, CD52 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 CD52. Human peripheral blood mononuclear cells (PBMCs) were stained with CD52 antibodies and with a suitable counterstaining. As a control, CD52 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 CD52. Human peripheral blood mononuclear cells (PBMCs) were stained with CD52 antibodies and with a suitable counterstaining. As a control, CD52 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 CD52 (REA164). 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 CD52 (REA164). 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 CD52 (REA164). 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 CD52 Antibody, anti-human, REAfinity™

Overview

Clone REA164 recognizes CD52, a glycosylphosphatidylinositol (GPI)-anchored, 25–29 kDa heavily glycosylated antigen. The CD52 core peptide is only 12 amino acids long and is linked to the surface via C-terminal GPI anchor. Expression of CD52 is found on lymphocytes, monocytes, macrophages, eosinophils, and in the male reproductive tract on the epithelial cells of the epididymis and seminal vesicle. CD52 is absent in immature spermatozoa in the efferent ducts but is acquired by the sperm membrane, progressively, as they pass through the epididymis proper, where CD52 is shed into the seminal plasma by the epididymis. Though the exact role of CD52 is unclear, antibodies against CD52 elicit antibody dependent cellular and complement mediated cytolysis of the CD52 expressing lymphocytes. Owing to this, such antibodies are often used to target malignant cells. CD52 expressed on sperms and on lymphocytes are basically different glycoforms of the same core peptide.
Additional information: Clone REA164 displays negligible binding to Fc receptors.

Alternative names

CDW52, CAMPATH-1

Detailed product information

Technical specifications

CloneREA164
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD52
Alternative names of antigenCDW52, CAMPATH-1
Molecular mass of antigen [kDa]1
Distribution of antigenlymphocytes
Entrez Gene ID1043
RRIDAB_2658646, AB_2658647, AB_2658648, AB_2658649, AB_2658650, AB_2658651, AB_2658652, AB_2658653, AB_2658654, AB_2658655, AB_2819508, AB_2819543

References for CD52 Antibody, anti-human, REAfinity™

Publications

  1. Ambrose, L. R. et al. (2009) Neutrophils express CD52 and exhibit complement-mediated lysis in the presence of alemtuzumab. Blood 114: 3052-3055
  2. Rawstron, A. C. et al. (2006) CD52 expression patterns in myeloma and the applicability of alemtuzumab therapy. Haematologica 91: 1577 -1578
  3. Yeung, C. H. et al. (1998) Epididymal secretion of CD52 as measured in human seminal plasma by a fluorescence immunoassay. Mol. Hum. Reprod. 4: 447-451

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