Invitrogen

CD95 (APO-1/Fas) Monoclonal Antibody (DX2), NovaFluor™ Yellow 610, eBioscience™

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Datasheet
Protocols
Questions & Answers
Datasheet
Protocols
Questions & Answers

Product Details

H084T03Y03-A

Applications
Tested Dilution
Publications

Flow Cytometry (Flow)

5 µL (0.6 µg)/test
-
Product Specifications

Species Reactivity

Dog, Human

Host/Isotype

Mouse / IgG1, kappa

Class

Monoclonal

Type

Antibody

Clone

DX2

Conjugate

NovaFluor™ Yellow 610 NovaFluor™ Yellow 610 NovaFluor™ Yellow 610

Excitation/Emission Max

551/614 nm View spectra spectra

Form

Liquid

Concentration

0.6 µg/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with BSA

Contains

0.09% sodium azide

Storage conditions

4°C, store in dark, DO NOT FREEZE!

RRID

AB_3666384

Product Specific Information

Description
The DX2 monoclonal antibody reacts with human CD95 (Fas, Apo-1), a 40-50 kDa member of the TNFR superfamily. DX2 does not block binding of EOS9.1, another antibody specific for human CD95. The DX2 monoclonal is reported to recognize dog/canine CD95.

This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.

Applications Tested
This DX2 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at (5 µL (0.6 µg)/test). A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.

Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.

Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.

CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 μL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 μL of CellBlox Plus Blocking Buffer per 100 μL of cell sample (10^3 to 10^8 cells).

NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more

Our internal testing shows that NovaFluor Yellow 610 non-specifically stains B cells in SJL mice. Non-specific staining has not been observed in BALB/c or C57BL/6 mice. Other strains have not been tested. See the Antibody Testing Data for an example of this strain-dependent difference.

Excitation: 551 nm; Emission: 614 nm; Laser: 561 nm (Yellow) Laser

Target Information

CD95, also known as Fas or APO-1, is a 46 kDa transmembrane glycoprotein that functions as a cell death receptor in the tumor necrosis factor receptor (TNFR) superfamily. It is expressed by a wide range of hematopoietic and non-hematopoietic cells, including monocytes, neutrophils, lymphocytes, and fibroblasts. CD95 also exists in a soluble form weighing approximately 26 kDa. The interaction of CD95 with its ligand, FasL, induces apoptosis, playing a crucial role in peripheral tolerance and anti-viral immune responses. Upon stimulation, CD95 leads to the aggregation of its intracellular Fas-associated death domains (FADD), formation of the death-inducing signaling complex (DISC), and activation of caspases. This cascade results in apoptosis. CD95 is also involved in activating NF-kappaB, MAPK3/ERK1, and MAPK8/JNK, and transduces proliferative signals in normal diploid fibroblasts and T cells. There are at least eight alternatively spliced transcript variants encoding seven distinct isoforms of CD95. Isoforms lacking the transmembrane domain may negatively regulate apoptosis mediated by the full-length isoform. The Fas/Fas ligand system is implicated in several human diseases, including AIDS, hepatitis, and cancer, highlighting its importance in immune regulation and disease pathology.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

How to use the Panel Builder

Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.

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Bioinformatics

Protein Aliases: Apo-1 antigen; APO-1 cell surface antigen; Apo-1 Fas; Apo1 antigen; apoptosis antigen 1; apoptosis signaling receptor FAS; Apoptosis-mediating surface antigen FAS; CD95; CD95 antigen; Fas (TNF receptor superfamily, member 6); Fas AMA; FASLG receptor; OTTHUMP00000020045; OTTHUMP00000020046; OTTHUMP00000020051; OTTHUMP00000059646; TNF receptor superfamily member 6; Tumor necrosis factor receptor superfamily member 6; tumor necrosis factor receptor superfamily, member 6

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Gene Aliases: ALPS1A; APO-1; APT1; CD95; FAS; FAS1; FASTM; TNFRSF6

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UniProt ID: (Human) Q14293

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Entrez Gene ID: (Dog) 486469, (Human) 355

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Function(s)
protease binding signal transducer activity receptor activity tumor necrosis factor-activated receptor activity protein binding kinase binding protein complex binding identical protein binding
Process(es)
ovarian follicle atresia immunoglobulin production renal system process protein complex assembly apoptotic process activation of cysteine-type endopeptidase activity involved in apoptotic process activation-induced cell death of T cells inflammatory cell apoptotic process inflammatory response immune response signal transduction spermatogenesis brain development aging circadian rhythm extrinsic apoptotic signaling pathway via death domain receptors response to toxic substance gene expression regulation of gene expression B cell mediated immunity telencephalon development dendrite regeneration positive regulation of protein homooligomerization response to lipopolysaccharide tumor necrosis factor-mediated signaling pathway regulation of cell proliferation response to drug ovulation cycle regulation of apoptotic process chordate embryonic development positive regulation of apoptotic process negative regulation of apoptotic process regulation of cysteine-type endopeptidase activity involved in apoptotic process positive regulation of MAPK cascade response to peptide hormone negative thymic T cell selection regulation of lymphocyte differentiation regulation of myeloid cell differentiation response to cycloheximide spleen development negative regulation of B cell activation protein homooligomerization response to glucocorticoid maternal process involved in female pregnancy positive regulation of lymphocyte apoptotic process cellular response to hydrogen peroxide response to growth factor cellular response to phenylalanine cellular response to mechanical stimulus cellular response to cobalt ion cellular response to lithium ion cellular response to glucose stimulus cellular response to interleukin-1 cellular response to estrogen stimulus cellular response to hyperoxia cellular response to hypoxia cellular response to hydrostatic pressure motor neuron apoptotic process apoptotic signaling pathway extrinsic apoptotic signaling pathway extrinsic apoptotic signaling pathway in absence of ligand hepatocyte apoptotic process activation of cysteine-type endopeptidase activity involved in apoptotic signaling pathway liver regeneration necroptotic signaling pathway regulation of extrinsic apoptotic signaling pathway via death domain receptors negative regulation of extrinsic apoptotic signaling pathway via death domain receptors response to fluoride positive regulation of extrinsic apoptotic signaling pathway in absence of ligand
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