Thermo Fisher Scientific Caspase 8 Monoclonal Antibody (1H10), eBioscience

Applications

Tested Dilution

Publications

Western Blot (WB)

1-5 µg/mL

-

Product Specifications

Species Reactivity

Human

Host/Isotype

Rat / IgG2a

Class

Monoclonal

Type

Antibody

Clone

1H10

Conjugate

Unconjugated Unconjugated Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4° C

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_1548833

Product Specific Information

Description: The monoclonal antibody 1H10 recognizes human Caspase-8 also known as FLICE and MACH, a member of the cysteine-requiring aspartate protease family. Caspase-8 is an upstream caspase responsible for Fas and TNF-alpha mediated apoptosis. After activation of the Fas or TNF receptors by their ligands, the adaptor molecule FADD binds to the receptor and recruits caspase-8 to the membrane. The formation of this complex induces the cleavage and activation of caspase-8. The active form of caspase-8, made of a heterotetramer of p18 and p10, is released in the cytoplasm and activates downstream apoptotic proteases (caspase-3, -4, -6, -7, -9 and -10). Several (> 8) isoforms have been identified: 3 different pro-forms called Caspase-8a (55kDA), Caspase-8b (53kDa), and Caspase-8L which is procaspase-8 with an additional N-terminal extension of 59 aa.

The 1H10 antibody recognizes the p55 and p53 proforms in addition to the p43 and p18 cleaved forms. Under reducing conditions, an additional band (possibly non-specific) around 35kDa may be present. Under nonreducing conditions, this band is absent.

Applications Reported: This 1H10 antibody has been reported for use in immunoblotting (WB).

Applications Tested: This 1H10 antibody has been tested by immunoblot analysis of cell lines. This can be used at 1-5 µg/mL It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Purity: Greater than 90%, as determined by SDS-PAGE.

Aggregation: Less than 10%, as determined by HPLC.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

Caspase 8 (CASP8) is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases play a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes composed of a pro-domain, 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. Caspase 8 is involved in the programmed cell death induced by Fas and various apoptotic stimuli. The N-terminal FADD-like death effector domain of Caspase 8 suggests that it may interact with Fas-interacting protein FADD. Caspase 8 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. Caspase 8 binds to the death effector domain (DED) of FADD through an analogous DED domain present in tandem in the pro-form of the Caspase 8 protein. Activated Caspase 8 then activates other downstream caspases including Caspase 9, thereby committing the cell to undergo apoptosis. In addition, Caspase 8 also reacts with Jurkat cells and Tonsil. Overexpression of Caspase 8 induces apoptosis, which can be blocked by inhibitors specific for the ICE family. Many alternatively spliced transcript variants encoding different isoforms have been described for Caspase 8, however, not all variants have had their full-length sequences determined.

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