Clone:
M-T271
Type of antibody:
Primary antibodies
Isotype:
mouse IgG1κ
Applications:
FC, MICS, IF, IHC
Alternative names:
S152, S152. LPFS2, T14, TNFRSF7, Tp55

Extended validation for CD27 Antibody, anti-human

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 M-T271
57703+
L128+
LG.3A10+
LG.7F9 (h/m/r)+
REA499+
O323++
Cells were incubated with an excess of purified unconjugated CD27 (M-T271) 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 CD27 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD27-PE (M-T271, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Fluorescence microscopy image of CD27 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD27-PE (M-T271, red) and counterstained with DRAQ5 (blue) as DNA stain.
Fluorescence microscopy image of CD27 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD27-PE (M-T271, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Overlay histogram showing flow cytometric analysis of CD27 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD27-PE, clone (M-T271). 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 CD27 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD27-PE, clone (M-T271). 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 CD27. Human peripheral blood mononuclear cells (PBMCs) were stained with CD27antibodies and with a suitable counterstaining. As a control, CD27 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 CD27. Human peripheral blood mononuclear cells (PBMCs) were stained with CD27antibodies and with a suitable counterstaining. As a control, CD27 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 CD27. Human peripheral blood mononuclear cells (PBMCs) were stained with CD27antibodies and with a suitable counterstaining. As a control, CD27 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 CD27. Human peripheral blood mononuclear cells (PBMCs) were stained with CD27antibodies and with a suitable counterstaining. As a control, CD27 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 CD27. Human peripheral blood mononuclear cells (PBMCs) were stained with CD27antibodies and with a suitable counterstaining. As a control, CD27 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 CD27. Human peripheral blood mononuclear cells (PBMCs) were stained with CD27antibodies and with a suitable counterstaining. As a control, CD27 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 CD27. Human peripheral blood mononuclear cells (PBMCs) were stained with CD27antibodies and with a suitable counterstaining. As a control, CD27 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 CD27. Human peripheral blood mononuclear cells (PBMCs) were stained with CD27antibodies and with a suitable counterstaining. As a control, CD27 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 CD27 (M-T271). 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 CD27 (M-T271). 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 CD27 (M-T271). 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 CD27 Antibody, anti-human

Overview

Clone M-T271 recognizes CD27, a member of the tumor necrosis factor receptor family. CD27 is expressed at differing levels on memory B cells, a fraction of plasma cells, and on naive and memory T cells, but is not expressed on naive B cells or effector T cells. It is also expressed on NK cells. CD27 therefore represents a useful marker to distinguish certain T and B cell subsets from each other.

Alternative names

S152, S152. LPFS2, T14, TNFRSF7, Tp55

Detailed product information

Technical specifications

CloneM-T271
Clonalitymonoclonal
Isotypemouse IgG1κ
Isotype controlIsotype Control Antibody, mouse IgG1
Hostmouse
Type of antibodyPrimary antibodies
Specieshuman, non-human primate
Cross-reactivity
rhesus monkey (
Macaca mulatta
)
,
cynomolgus monkey (
Macaca fascicularis
)
AntigenCD27
Alternative names of antigenS152, S152. LPFS2, T14, TNFRSF7, Tp55
Molecular mass of antigen [kDa]27
Distribution of antigenB cells, NK cells, red blood cells, T cells, plasma cells, thymocytes
Entrez Gene ID939
RRIDAB_2733629, AB_2751225, AB_2751160, AB_2751222, AB_2751157, AB_2751220, AB_2751154, AB_2751224, AB_2751159, AB_2751226, AB_2751161, AB_2751223, AB_2751158, AB_2751221, AB_2751155, AB_2784097, AB_2784096, AB_2751156, AB_2733628

Resources for CD27 Antibody, anti-human

Certificates

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