Anti-Fas, clone SM1/1 (mouse monoclonal)

Species Reactivity	Key Applications	Host	Format	Antibody Type
H	FC, FUNC	M	Purified	Monoclonal Antibody

Species Reactivity

Key Applications

Host

Format

Antibody Type

FC, FUNC

Purified

Monoclonal Antibody

Description
Catalogue Number	MAB3061
Replaces	CBL527B
Brand Family	Chemicon®
Trade Name	Chemicon
Description	Anti-Fas Antibody, clone SM1/1
Alternate Names	CD95 Apo-1

Description

Catalogue Number

MAB3061

Replaces

CBL527B

Brand Family

Chemicon®

Trade Name

Chemicon

Description

Anti-Fas Antibody, clone SM1/1

Alternate Names

CD95
Apo-1

Product Information
Format	Purified
Presentation	Purified IgG provided in PBS, no preservatives.

Product Information

Format

Purified

Presentation

Purified IgG provided in PBS, no preservatives.

Applications
Application	This Anti-Fas Antibody, clone SM1/1 is validated for use in FC, FUNC for the detection of Fas.
Key Applications	Flow Cytometry Affects Function
Application Notes	Flow cytometry: 1-10 μg/mL, with incubation for 30 minutes at 4°C. Daudi or L929 cells may be used as negative controls. Induction of apoptosis with SM1/1: SM1/1 induces apoptosis best when used together with a 10 molar excess of crosslinking anti-mouse IgG. The amount of antibody needed will vary depending upon the cell line used and the age of the cell and their growing conditions; best results are achieved when cells are less than 70% confluent and relatively young passage numbers, and note not all cells what express CD95 can be induced with SM1/1, the reasons why are unclear. On Jurkat cells 100-500ng/mL of SM1/1 in the presence of 10X excess of goat anti-mouse IgG will produce ~50% kill as measured by MTT assay; without crosslinking, little or no killing will be observed. On SKw6.4 cells 100-500ng/mL of SM1/1 in the presence of 10X excess goat anti-mouse IgG will produce ~50% kill as measured by MTT; without crosslinking ~20% or less of the cells will be induced. On L/F15 cells 100-500ng/mL of SM1/1 with or without crosslinking IgG ~ 50% of the cells will be induced to undergo apoptosis as measured by MTT assay. In all cases it is important to remove excess SM1/1 before adding the secondary crosslinking antibody; Antibody additions can be either at 4C, or 37C; at 37C primary antibody should be incubated no longer than two hours prior to secondary crosslinker addition. Crosslinking antibody is typically incubated overnight, although shorter times may yield acceptable results. As controls, cells should be incubated with an unrelated irrelevant isotype matched IgG2a antibody or with the crosslinking antibody alone. SM1/1 monoclonal characterization is first reported in Trauth, BC et al Science 1989 245:301-5. Optimal working dilutions must be determined by end user.

Applications

Application

This Anti-Fas Antibody, clone SM1/1 is validated for use in FC, FUNC for the detection of Fas.

Key Applications

Flow Cytometry
Affects Function

Application Notes

Flow cytometry: 1-10 μg/mL, with incubation for 30 minutes at 4°C. Daudi or L929 cells may be used as negative controls.

Induction of apoptosis with SM1/1:

SM1/1 induces apoptosis best when used together with a 10 molar excess of crosslinking anti-mouse IgG. The amount of antibody needed will vary depending upon the cell line used and the age of the cell and their growing conditions; best results are achieved when cells are less than 70% confluent and relatively young passage numbers, and note not all cells what express CD95 can be induced with SM1/1, the reasons why are unclear.

On Jurkat cells 100-500ng/mL of SM1/1 in the presence of 10X excess of goat anti-mouse IgG will produce ~50% kill as measured by MTT assay; without crosslinking, little or no killing will be observed.

On SKw6.4 cells 100-500ng/mL of SM1/1 in the presence of 10X excess goat anti-mouse IgG will produce ~50% kill as measured by MTT; without crosslinking ~20% or less of the cells will be induced.

On L/F15 cells 100-500ng/mL of SM1/1 with or without crosslinking IgG ~ 50% of the cells will be induced to undergo apoptosis as measured by MTT assay.

In all cases it is important to remove excess SM1/1 before adding the secondary crosslinking antibody;

Antibody additions can be either at 4C, or 37C; at 37C primary antibody should be incubated no longer than two hours prior to secondary crosslinker addition. Crosslinking antibody is typically incubated overnight, although shorter times may yield acceptable results.

As controls, cells should be incubated with an unrelated irrelevant isotype matched IgG2a antibody or with the crosslinking antibody alone.

SM1/1 monoclonal characterization is first reported in Trauth, BC et al Science 1989 245:301-5.

Optimal working dilutions must be determined by end user.

Biological Information
Clone	SM1/1
Concentration	Please refer to the Certificate of Analysis for the lot-specific concentration.
Host	Mouse
Specificity	Specifically recognizes Fas [CD95/APO-1]
Isotype	IgG2a
Species Reactivity	Human
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NM_000043.3 NM_152871.1 NM_152872.1 NM_152873.1 NM_152874.1 NM_152875.1 NM_152876.1 NM_152877.1
Entrez Gene Summary	The protein encoded by this gene is a member of the TNF-receptor superfamily. This receptor contains a death domain. It has been shown to play a central role in the physiological regulation of programmed cell death, and has been implicated in the pathogenesis of various malignancies and diseases of the immune system. The interaction of this receptor with its ligand allows the formation of a death-inducing signaling complex that includes Fas-associated death domain protein (FADD), caspase 8, and caspase 10. The autoproteolytic processing of the caspases in the complex triggers a downstream caspase cascade, and leads to apoptosis. This receptor has been also shown to activate NF-kappaB, MAPK3/ERK1, and MAPK8/JNK, and is found to be involved in transducing the proliferating signals in normal diploid fibroblast and T cells. At least eight alternatively spliced transcript variants encoding seven distinct isoforms have been described. The isoforms lacking the transmembrane domain may negatively regulate the apoptosis mediated by the full length isoform.
Gene Symbol	FAS APT1 CD95 FASTM NMP35 FAS1 APO-1 ALPS1A FAIM2 OTTHUMP00000059646 TNFRSF6 LFG
UniProt Number	P25445
UniProt Summary	FUNCTION: SwissProt: P25445 # Receptor for TNFSF6/FASLG. The adapter molecule FADD recruits caspase-8 to the activated receptor. The resulting death- inducing signaling complex (DISC) performs caspase-8 proteolytic activation which initiates the subsequent cascade of caspases (aspartate-specific cysteine proteases) mediating apoptosis. FAS- mediated apoptosis may have a role in the induction of peripheral tolerance, in the antigen-stimulated suicide of mature T-cells, or both. The secreted isoforms 2 to 6 block apoptosis (in vitro). SIZE: 335 amino acids; 37732 Da SUBUNIT: Binds DAXX. Interacts with HIPK3. Part of a complex containing HIPK3 and FADD (By similarity). Binds RIPK1 and FAIM2. Interacts with BRE. SUBCELLULAR LOCATION: Isoform 1: Cell membrane; Single-pass type I membrane protein. & Isoform 2: Secreted. & Isoform 3: Secreted. & Isoform 4: Secreted. & Isoform 5: Secreted. & Isoform 6: Secreted. TISSUE SPECIFICITY: Isoform 1 and isoform 6 are expressed at equal levels in resting peripheral blood mononuclear cells. After activation there is an increase in isoform 1 and decrease in the levels of isoform 6. DOMAIN: SwissProt: P25445 Contains a death domain involved in the binding of FADD, and maybe to other cytosolic adapter proteins. DISEASE: SwissProt: P25445 # Defects in FAS are the cause of autoimmune lymphoproliferative syndrome type 1A (ALPS1A) [MIM:601859]; also known as Canale-Smith syndrome (CSS). ALPS is a childhood syndrome involving hemolytic anemia and thrombocytopenia with massive lymphadenopathy and splenomegaly. SIMILARITY: Contains 1 death domain. & Contains 3 TNFR-Cys repeats.

Biological Information

Clone

SM1/1

Concentration

Please refer to the Certificate of Analysis for the lot-specific concentration.

Host

Mouse

Specificity

Specifically recognizes Fas [CD95/APO-1]

Isotype

IgG2a

Species Reactivity

Human

Antibody Type

Monoclonal Antibody

Entrez Gene Number

Entrez Gene Summary

The protein encoded by this gene is a member of the TNF-receptor superfamily. This receptor contains a death domain. It has been shown to play a central role in the physiological regulation of programmed cell death, and has been implicated in the pathogenesis of various malignancies and diseases of the immune system. The interaction of this receptor with its ligand allows the formation of a death-inducing signaling complex that includes Fas-associated death domain protein (FADD), caspase 8, and caspase 10. The autoproteolytic processing of the caspases in the complex triggers a downstream caspase cascade, and leads to apoptosis. This receptor has been also shown to activate NF-kappaB, MAPK3/ERK1, and MAPK8/JNK, and is found to be involved in transducing the proliferating signals in normal diploid fibroblast and T cells. At least eight alternatively spliced transcript variants encoding seven distinct isoforms have been described. The isoforms lacking the transmembrane domain may negatively regulate the apoptosis mediated by the full length isoform.

Gene Symbol

FAS
APT1
CD95
FASTM
NMP35
FAS1
APO-1
ALPS1A
FAIM2
OTTHUMP00000059646
TNFRSF6
LFG

UniProt Number

P25445

UniProt Summary

FUNCTION: SwissProt: P25445 # Receptor for TNFSF6/FASLG. The adapter molecule FADD recruits caspase-8 to the activated receptor. The resulting death- inducing signaling complex (DISC) performs caspase-8 proteolytic activation which initiates the subsequent cascade of caspases (aspartate-specific cysteine proteases) mediating apoptosis. FAS- mediated apoptosis may have a role in the induction of peripheral tolerance, in the antigen-stimulated suicide of mature T-cells, or both. The secreted isoforms 2 to 6 block apoptosis (in vitro).
SIZE: 335 amino acids; 37732 Da
SUBUNIT: Binds DAXX. Interacts with HIPK3. Part of a complex containing HIPK3 and FADD (By similarity). Binds RIPK1 and FAIM2. Interacts with BRE.
SUBCELLULAR LOCATION: Isoform 1: Cell membrane; Single-pass type I membrane protein. & Isoform 2: Secreted. & Isoform 3: Secreted. & Isoform 4: Secreted. & Isoform 5: Secreted. & Isoform 6: Secreted.
TISSUE SPECIFICITY: Isoform 1 and isoform 6 are expressed at equal levels in resting peripheral blood mononuclear cells. After activation there is an increase in isoform 1 and decrease in the levels of isoform 6.
DOMAIN: SwissProt: P25445 Contains a death domain involved in the binding of FADD, and maybe to other cytosolic adapter proteins.
DISEASE: SwissProt: P25445 # Defects in FAS are the cause of autoimmune lymphoproliferative syndrome type 1A (ALPS1A) [MIM:601859]; also known as Canale-Smith syndrome (CSS). ALPS is a childhood syndrome involving hemolytic anemia and thrombocytopenia with massive lymphadenopathy and splenomegaly.
SIMILARITY: Contains 1 death domain. & Contains 3 TNFR-Cys repeats.

Product Usage Statements
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Maintain at 2-8°C in undiluted aliquots. After opening, aliquot and freeze at -20°C. Avoid repeated freeze/thaw cycles.

Storage and Shipping Information

Storage Conditions

Maintain at 2-8°C in undiluted aliquots. After opening, aliquot and freeze at -20°C. Avoid repeated freeze/thaw cycles.

Packaging Information
Material Size	100 µg

Packaging Information

Material Size

100 µg

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

Anti-Fas, clone SM1/1 (mouse monoclonal)

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