Clone:
REA453
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Alternative names:
FAS, APO-1, Apt1, Tnfrsf6, TNFR6, lpr

Extended validation for CD95 (FAS) Antibody, anti-mouse, 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 REA453
Jo2++
SA367H8-
Cells were incubated with an excess of purified unconjugated CD95 (FAS) (REA453) 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 CD95 (FAS) (specificity from PIM). Thymocytes from BALB/c mice were stained with CD95 (FAS) (specificity from PIM) antibodies and with a suitable counterstaining. As a control, CD95 (FAS) 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. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD95 (FAS) (specificity from PIM). Thymocytes from BALB/c mice were stained with CD95 (FAS) (specificity from PIM) antibodies and with a suitable counterstaining. As a control, CD95 (FAS) 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. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD95 (FAS) (specificity from PIM). Thymocytes from BALB/c mice were stained with CD95 (FAS) (specificity from PIM) antibodies and with a suitable counterstaining. As a control, CD95 (FAS) 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. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD95 (FAS) (specificity from PIM). Thymocytes from BALB/c mice were stained with CD95 (FAS) (specificity from PIM) antibodies and with a suitable counterstaining. As a control, CD95 (FAS) 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. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD95 (FAS) (specificity from PIM). Thymocytes from BALB/c mice were stained with CD95 (FAS) (specificity from PIM) antibodies and with a suitable counterstaining. As a control, CD95 (FAS) 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. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for CD95 (FAS) (specificity from PIM). Thymocytes from BALB/c mice were stained with CD95 (FAS) (specificity from PIM) antibodies and with a suitable counterstaining. As a control, CD95 (FAS) 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. 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 CD95 (FAS) (REA453). 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 CD95 (FAS) (REA453). 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 CD95 (FAS) (REA453). 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 CD95 (FAS) Antibody, anti-mouse, REAfinity™

Overview

Clone REA453 recognizes the mouse CD95 antigen, a single-pass type I membrane protein which is also known as FAS or Apo-1. CD95 is a member of the tumor necrosis factor receptor superfamily and is found on the surface of many normal and neoplastically transformed cells. It is an apoptosis-inducing death receptor that regulates tissue homeostasis mainly in the immune system through induction of apoptosis by binding of its ligand CD95L (FASL/Apo-1L). CD95 and CD95L are up-regulated on lymphocytes upon activation and are known to play a key role in the regulation of an inflammatory response. CD95 also plays an apoptosis-independent role in non-immune cells and it has been implicated in cancer cell growth, migration, and tumor progression. During cancer progression CD95 is frequently down regulated or cells are rendered apoptosis resistant raising the possibility that loss of CD95 is part of a mechanism for tumor evasion.
Additional information: Clone REA453 displays negligible binding to Fc receptors.

Alternative names

FAS, APO-1, Apt1, Tnfrsf6, TNFR6, lpr

Detailed product information

Technical specifications

CloneREA453
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD95 (FAS)
Alternative names of antigenFAS, APO-1, Apt1, Tnfrsf6, TNFR6, lpr
Molecular mass of antigen [kDa]35
Distribution of antigenlymphocytes
Entrez Gene ID14102
RRIDAB_2751726, AB_2801758, AB_2659654, AB_2659655, AB_2751784

References for CD95 (FAS) Antibody, anti-mouse, REAfinity™

Publications

  1. Watanabe-Fukunaga, R. et al. (1992) The cDNA structure, expression, and chromosomal assignment of the mouse Fas antigen. J. Immunol. 148(4): 1274-1279
  2. Chen, L. et al. (2010) CD95 promotes tumour growth. Nature 465(7297): 492-496
  3. Hadji, A. et al. (2014) Death induced by CD95 or CD95 ligand elimination. Cell Rep 7(1): 208-222

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