Clone:
LG.3A10
Type of antibody:
Primary antibodies
Isotype:
hamster IgG
Applications:
FC, MICS, IF, IHC, ICC
Alternative names:
S152, S152. LPFS2, T14, TNFRSF7, Tp55

Extended validation for CD27 Antibody, anti-human/mouse

Specificity

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
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Fluorescence microscopy image of CD27 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD27-PE (LG.3A10, red) and counterstained with DRAQ5 (blue) as DNA stain.
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Fluorescence microscopy image of CD27 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD27-PE (LG.3A10, 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 (LG.3A10, 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 (LG.3A10). 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 (LG.3A10). 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
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Comparison of staining pattern on non-fixed and fixed cells using CD27 (LG.3A10). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
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Comparison of staining pattern on non-fixed and fixed cells using CD27 (LG.3A10). 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 (LG.3A10). 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/mouse

Overview

Clone LG.3A10 recognizes the human and murine CD27 antigen, a member of the tumor necrosis factor receptor family. CD27 is expressed on subsets of B, T, and NK cells. CD27 binds to CD70. This interaction regulates cell activation.

Alternative names

S152, S152. LPFS2, T14, TNFRSF7, Tp55

Detailed product information

Technical specifications

CloneLG.3A10
Clonalitymonoclonal
Isotypehamster IgG
Hosthamster
Type of antibodyPrimary antibodies
Specieshuman, mouse, rat
Cross-reactivityrat
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_2656819, AB_2656820, AB_2656821, AB_2656822, AB_2656823, AB_2656824, AB_2656825, AB_2656818

References for CD27 Antibody, anti-human/mouse

Publications

  1. Gravestein, L. A. et al. (1995) Novel mAbs reveal potent co-stimulatory activity of murine CD27. Int. Immunol. 7: 551-557

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