Clone:
DX2
Type of antibody:
Primary antibodies, Functional-grade antibodies
Isotype:
mouse IgG1κ
Applications:
FC, MICS, IF, IHC, ICC, FA, MC
Alternative names:
FAS, ALPS1A, APO-1, Apt1, FAS1, FASTM, Tnfrsf6

Extended validation for CD95 (FAS) 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 DX2
REA738++
EOS9.1-
LT95++
UB2n/a
B-10n/a
4F8H6-
SM1/23++
Cells were incubated with an excess of purified unconjugated CD95 (FAS) (DX2) 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 CD95 (FAS) knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD95 (FAS)-PE (DX2, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Fluorescence microscopy image of CD95 (FAS) knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD95 (FAS)-PE (DX2, red) and counterstained with DRAQ5 (blue) as DNA stain.
Fluorescence microscopy image of CD95 (FAS) knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD95 (FAS)-PE (DX2, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Overlay histogram showing flow cytometric analysis of CD95 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD95-PE, clone (DX2). 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 CD95 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD95-PE, clone (DX2). 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.

Specifications for CD95 (FAS) Antibody, anti-human

Overview

CD95, also known as FAS or Apo-1, is a member of the tumor necrosis factor receptor superfamily (TNFR) and is found on the surface of many normal and neoplastically transformed cells. Its ligand, CD95L (FASL/Apo-1L), is able to induce apoptosis in CD95-expressing cells upon binding. 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: Juxtocrine “fratricide” of neighbouring lymphocytes via mutual CD95 and CD95L expression helps to terminate immune responses, while apoptosis of pro-inflammatory cells via CD95 helps maintain immune privilege in sites such as the eye, where CD95L is found to be expressed in the retina and cornea. Cross-linking of CD95 receptors by DX2 monoclonal antibody has been described to induce apoptosis in certain target cells.

Alternative names

FAS, ALPS1A, APO-1, Apt1, FAS1, FASTM, Tnfrsf6

Detailed product information

Technical specifications

CloneDX2
Clonalitymonoclonal
Isotypemouse IgG1κ
Isotype controlIsotype Control Antibody, mouse IgG1
Hostmouse
Type of antibodyPrimary antibodies, Functional-grade antibodies
Specieshuman, non-human primate
Cross-reactivity
rhesus monkey (
Macaca mulatta
)
,
cynomolgus monkey (
Macaca fascicularis
)
AntigenCD95 (FAS)
Alternative names of antigenFAS, ALPS1A, APO-1, Apt1, FAS1, FASTM, Tnfrsf6
Molecular mass of antigen [kDa]35
Distribution of antigenB cells, epithelial cells, fibroblasts, monocytes, T cells, neutrophils, thymocytes, heart, kidney, liver, lung, ovary
Entrez Gene ID355
RRIDAB_2751411, AB_2728048, AB_2728025, AB_2811577, AB_2811546, AB_2811589, AB_2811569, AB_2819597, AB_2819582, AB_2751041, AB_2660559, AB_871701, AB_2751429

References for CD95 (FAS) Antibody, anti-human

Publications

  1. Pitcher, C. J. et al. (2002)
    Development and homeostasis of T cell memory in
    rhesus macaque
    .
    J. Immunol. 168: 29-43
  2. Griffith, T. et al. (1995) Fas ligand-induced apoptosis as a mechanism of immune privilege. Science 270: 1189-1192
  3. Itoh, N. et al. (1991) The polypeptide encoded by the cDNA for human cell surface antigen Fas can mediate apoptosis. Cell 66: 233-243
  4. Komada, Y. et al. (1999) Epitopes and functional responses defined by a panel of anti-Fas (CD95) monoclonal antibodies. Hybridoma 18: 391-398
  5. Lynch, D. et al. (1995) Fas and FasL in the homeostatic regulation of immune responses. Immunol. Today 16: 569-574

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