Anti-Caspase-8 (active form p18 subunit), clone 2B12.1 (mouse monoclonal)

Species Reactivity	Key Applications	Host	Format	Antibody Type
H	WB, ICC	M	Purified	Monoclonal Antibody

Species Reactivity

Key Applications

Host

Format

Antibody Type

WB, ICC

Purified

Monoclonal Antibody

Description
Catalogue Number	MAB10754
Replaces	04-573; 04-574
Description	Anti-Caspase-8 (active form p18 subunit) Antibody, clone 2B12.1
Alternate Names	caspase 8, apoptosis-related cysteine peptidase caspase-8 caspase 8, apoptosis-related cysteine protease MACH-beta-1/2/3/4 protein Apoptotic protease Mch-5 Apoptotic cysteine protease MACH-alpha-1/2/3 protein FADD-homologous ICE/ced-3-like protease MORT1-associated ced-3 homolog ICE-like apoptotic protease 5 FADD-like ICE
Background Information	Caspase-8 (MACH/FLICE/Mch5) is a member of a family of cysteine-requiring aspartate proteases that participate in the intracellular signaling cascade leading to apoptosis, or programmed cell death. Caspase-8 is recruited to death-inducing signaling complex (DISC), CD95 (Fas/APO-1) and tumor necrosis factor receptor 1 (TNFR1) and is subsequently activated by proteolytic cleavage into two residues, p10 and p18. Active caspase-8 or the p18 subunit, is the first in the caspase signaling cascade, and it signals downstream caspases to eventually induce apoptosis.

Description

Catalogue Number

MAB10754

Replaces

04-573; 04-574

Description

Anti-Caspase-8 (active form p18 subunit) Antibody, clone 2B12.1

Alternate Names

caspase 8, apoptosis-related cysteine peptidase
caspase-8
caspase 8, apoptosis-related cysteine protease
MACH-beta-1/2/3/4 protein
Apoptotic protease Mch-5
Apoptotic cysteine protease
MACH-alpha-1/2/3 protein
FADD-homologous ICE/ced-3-like protease
MORT1-associated ced-3 homolog
ICE-like apoptotic protease 5
FADD-like ICE

Background Information

Caspase-8 (MACH/FLICE/Mch5) is a member of a family of cysteine-requiring aspartate proteases that participate in the intracellular signaling cascade leading to apoptosis, or programmed cell death. Caspase-8 is recruited to death-inducing signaling complex (DISC), CD95 (Fas/APO-1) and tumor necrosis factor receptor 1 (TNFR1) and is subsequently activated by proteolytic cleavage into two residues, p10 and p18. Active caspase-8 or the p18 subunit, is the first in the caspase signaling cascade, and it signals downstream caspases to eventually induce apoptosis.

Product Information
Format	Purified
Control	Camptothecin-treated and untreated Jurkat lysates
Presentation	Purified mouse monoclonal IgMκ in buffer containing PBS with 0.05% sodium azide.

Product Information

Format

Purified

Control

Camptothecin-treated and untreated Jurkat lysates

Presentation

Purified mouse monoclonal IgMκ in buffer containing PBS with 0.05% sodium azide.

Applications
Application	Detect Caspase-8 (active form p18 subunit) using this Anti-Caspase-8 (active form p18 subunit) Antibody, clone 2B12.1 validated for use in WB & IC.
Key Applications	Western Blotting Immunocytochemistry
Application Notes	Immunocytochemistry Analysis: A 1:500 dilution from a previous lot detected active Caspase-8 (p18 subunit) in etoposide-treated HeLa cells.

Applications

Application

Detect Caspase-8 (active form p18 subunit) using this Anti-Caspase-8 (active form p18 subunit) Antibody, clone 2B12.1 validated for use in WB & IC.

Key Applications

Western Blotting
Immunocytochemistry

Application Notes

Immunocytochemistry Analysis: A 1:500 dilution from a previous lot detected active Caspase-8 (p18 subunit) in etoposide-treated HeLa cells.

Biological Information
Immunogen	KLH-conjugated linear peptide corresponding to the active form of human Caspase-8.
Clone	2B12.1
Concentration	Please refer to the Certificate of Analysis for the lot-specific concentration.
Host	Mouse
Specificity	This antibody recognizes the active form (p18 subunit) of Caspase-8.
Isotype	IgMκ
Species Reactivity	Human
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NP_001073593
Entrez Gene Summary	This gene encodes a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes composed of a prodomain, a large protease subunit, and a small protease subunit. Activation of caspases requires proteolytic processing at conserved internal aspartic residues to generate a heterodimeric enzyme consisting of the large and small subunits. This protein is involved in the programmed cell death induced by Fas and various apoptotic stimuli. The N-terminal FADD-like death effector domain of this protein suggests that it may interact with Fas-interacting protein FADD. This protein was detected in the insoluble fraction of the affected brain region from Huntington disease patients but not in those from normal controls, which implicated the role in neurodegenerative diseases. Many alternatively spliced transcript variants encoding different isoforms have been described, although not all variants have had their full-length sequences determined. [provided by RefSeq].
Gene Symbol	CASP8 MCH5 CASP-8 MACH ALPS2B FLICE procaspase-8 CAP4
Purification Method	Ion Exchange
UniProt Number	Q14790
UniProt Summary	FUNCTION: Most upstream protease of the activation cascade of caspases responsible for the TNFRSF6/FAS mediated and TNFRSF1A induced cell death. Binding to the adapter molecule FADD recruits it to either receptor. The resulting aggregate called death- inducing signaling complex (DISC) performs CASP8 proteolytic activation. The active dimeric enzyme is then liberated from the DISC and free to activate downstream apoptotic proteases. Proteolytic fragments of the N-terminal propeptide (termed CAP3, CAP5 and CAP6) are likely retained in the DISC. Cleaves and activates CASP3, CASP4, CASP6, CASP7, CASP9 and CASP10. May participate in the GZMB apoptotic pathways. Cleaves ADPRT. Hydrolyzes the small-molecule substrate, Ac-Asp-Glu-Val-Asp- -AMC. Likely target for the cowpox virus CRMA death inhibitory protein. Isoforms 5, 6, 7 and 8 lack the catalytic site and may interfere with the pro-apoptotic activity of the complex. SIZE: 479 amino acids; 55391 Da SUBUNIT: Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 18 kDa (p18) and a 10 kDa (p10) subunit. Interacts with FADD, CFLAR and PEA15. Isoform 9 interacts at the endoplasmic reticulum with a complex containing BCAP31, BAP29, BCL2 and/or BCL2L1. SUBCELLULAR LOCATION: Cytoplasm. TISSUE SPECIFICITY: Isoforms 1, 5 and 7 are expressed in a wide variety of tissues. Highest expression in peripheral blood leukocytes, spleen, thymus, and liver. Barely detectable in brain, testis, and skeletal muscle.DOMAIN:SwissProt: Q14790 Isoform 9 contains a N-terminal extension that is required for interaction with the BCAP31 complex. PTM: Generation of the subunits requires association with the death-inducing signaling complex (DISC), whereas additional processing is likely due to the autocatalytic activity of the activated protease. GZMB and CASP10 can be involved in these processing events. & Phosphorylated upon DNA damage, probably by ATM or ATR. DISEASE: Defects in CASP8 are the cause of caspase-8 deficiency (CASP8D) [MIM:607271]. CASP8D is a disorder resembling autoimmune lymphoproliferative syndrome (ALPS). It is characterized by lymphadenopathy, splenomegaly, and defective CD95-induced apoptosis of peripheral blood lymphocytes (PBLs). It leads to defects in activation of T-lymphocytes, B-lymphocytes, and natural killer cells leading to immunodeficiency characterized by recurrent sinopulmonary and herpes simplex virus infections and poor responses to immunization. SIMILARITY: Belongs to the peptidase C14 family. & Contains 2 DED (death effector) domains.
Molecular Weight	~18 kDa observed

Biological Information

Immunogen

KLH-conjugated linear peptide corresponding to the active form of human Caspase-8.

Clone

2B12.1

Concentration

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

Host

Mouse

Specificity

This antibody recognizes the active form (p18 subunit) of Caspase-8.

Isotype

IgMκ

Species Reactivity

Human

Antibody Type

Monoclonal Antibody

Entrez Gene Number

NP_001073593

Entrez Gene Summary

This gene encodes a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes composed of a prodomain, a large protease subunit, and a small protease subunit. Activation of caspases requires proteolytic processing at conserved internal aspartic residues to generate a heterodimeric enzyme consisting of the large and small subunits. This protein is involved in the programmed cell death induced by Fas and various apoptotic stimuli. The N-terminal FADD-like death effector domain of this protein suggests that it may interact with Fas-interacting protein FADD. This protein was detected in the insoluble fraction of the affected brain region from Huntington disease patients but not in those from normal controls, which implicated the role in neurodegenerative diseases. Many alternatively spliced transcript variants encoding different isoforms have been described, although not all variants have had their full-length sequences determined. [provided by RefSeq].

Gene Symbol

CASP8
MCH5
CASP-8
MACH
ALPS2B
FLICE
procaspase-8
CAP4

Purification Method

Ion Exchange

UniProt Number

Q14790

UniProt Summary

FUNCTION: Most upstream protease of the activation cascade of caspases responsible for the TNFRSF6/FAS mediated and TNFRSF1A induced cell death. Binding to the adapter molecule FADD recruits it to either receptor. The resulting aggregate called death- inducing signaling complex (DISC) performs CASP8 proteolytic activation. The active dimeric enzyme is then liberated from the DISC and free to activate downstream apoptotic proteases. Proteolytic fragments of the N-terminal propeptide (termed CAP3, CAP5 and CAP6) are likely retained in the DISC. Cleaves and activates CASP3, CASP4, CASP6, CASP7, CASP9 and CASP10. May participate in the GZMB apoptotic pathways. Cleaves ADPRT. Hydrolyzes the small-molecule substrate, Ac-Asp-Glu-Val-Asp- -AMC. Likely target for the cowpox virus CRMA death inhibitory protein. Isoforms 5, 6, 7 and 8 lack the catalytic site and may interfere with the pro-apoptotic activity of the complex.

SIZE: 479 amino acids; 55391 Da

SUBUNIT: Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 18 kDa (p18) and a 10 kDa (p10) subunit. Interacts with FADD, CFLAR and PEA15. Isoform 9 interacts at the endoplasmic reticulum with a complex containing BCAP31, BAP29, BCL2 and/or BCL2L1.

SUBCELLULAR LOCATION: Cytoplasm.

TISSUE SPECIFICITY: Isoforms 1, 5 and 7 are expressed in a wide variety of tissues. Highest expression in peripheral blood leukocytes, spleen, thymus, and liver. Barely detectable in brain, testis, and skeletal muscle.DOMAIN:SwissProt: Q14790 Isoform 9 contains a N-terminal extension that is required for interaction with the BCAP31 complex.

PTM: Generation of the subunits requires association with the death-inducing signaling complex (DISC), whereas additional processing is likely due to the autocatalytic activity of the activated protease. GZMB and CASP10 can be involved in these processing events. & Phosphorylated upon DNA damage, probably by ATM or ATR.

DISEASE: Defects in CASP8 are the cause of caspase-8 deficiency (CASP8D) [MIM:607271]. CASP8D is a disorder resembling autoimmune lymphoproliferative syndrome (ALPS). It is characterized by lymphadenopathy, splenomegaly, and defective CD95-induced apoptosis of peripheral blood lymphocytes (PBLs). It leads to defects in activation of T-lymphocytes, B-lymphocytes, and natural killer cells leading to immunodeficiency characterized by recurrent sinopulmonary and herpes simplex virus infections and poor responses to immunization.

SIMILARITY: Belongs to the peptidase C14 family. & Contains 2 DED (death effector) domains.

Molecular Weight

~18 kDa observed

Product Usage Statements
Quality Assurance	Evaluated by Western Blot in camptothecin-treated and untreated Jurkat lysates. Western Blot Analysis: 2 µg/mL of this antibody detected the active form of Caspase-8 (p18 subunit) in 10 µg of camptothecin-treated Jurkat lysates. Untreated jurkat lysate exhibited no detection of active Caspase-8.
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

Quality Assurance

Evaluated by Western Blot in camptothecin-treated and untreated Jurkat lysates.

Western Blot Analysis: 2 µg/mL of this antibody detected the active form of Caspase-8 (p18 subunit) in 10 µg of camptothecin-treated Jurkat lysates. Untreated jurkat lysate exhibited no detection of active Caspase-8.

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	Stable for 1 year at 2-8°C from date of receipt.

Storage and Shipping Information

Storage Conditions

Stable for 1 year at 2-8°C from date of receipt.

Packaging Information
Material Size	200 µg

Packaging Information

Material Size

200 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-Caspase-8 (active form p18 subunit), clone 2B12.1 (mouse monoclonal)

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