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Caspase-8 Overexpression Lysate

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Western Blot: Caspase 8 Overexpression Lysate (Adult Normal) [NBL1-08717] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Caspase 8.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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Caspase-8 Overexpression Lysate Summary

Description

Caspase-8 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC216990

Accession#: NM_033355

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
Gene
CASP8

Applications/Dilutions

Dilutions
  • Western Blot
Application Notes
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
Theoretical MW
55.2 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

Storage
Store at -80C. Avoid freeze-thaw cycles.
Buffer
RIPA buffer

Lysate Details for Array

Type
Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for Caspase-8 Overexpression Lysate

  • AIS
  • androgen receptor
  • CASP8
  • Caspase8
  • Caspase-8
  • DHTRTFM
  • Dihydrotestosterone receptorHYSP1
  • HUMARA
  • Mch5
  • NR3C4KD
  • Nuclear receptor subfamily 3 group C member 4
  • SMAX1SBMA
  • spinal and bulbar muscular atrophy

Background

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]. Transcript Variant: This variant (B) includes different segments in its 5' UTR and lacks a 5' coding region segment, compared to variant G. This results in translation at a downstream start codon, and the encoded protein (isoform B) has a shorter N-terminus when it is compared to isoform G. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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Publications for Caspase-8 Lysate (NBL1-08717) (0)

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Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol

FAQs for Caspase-8 Lysate (NBL1-08717). (Showing 1 - 1 of 1 FAQ).

  1. May we ask the MW difference between the active form and the proform of Caspase 8 while performing WB?
    • In treated cells induced to undergo apoptosis, caspase-8 migrates as a 55/53 kDa (pro-form), 41/42 kDa (a cleaved/active or intermediate form), and 18 kDa (active form). The proform (55/53 kDa) is still seen in treated cells because not all cells undergo apoptosis at once.

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Research Areas for Caspase-8 Lysate (NBL1-08717)

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Blogs on Caspase-8.

The NLRP3 Inflammasome: Macrophage Activator & Pathology Driver
By Victoria Osinski, PhDWhat is the NLRP3 Inflammasome?With its critical role in innate immunity, the nucleotide-binding oligomerization domain-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome ...  Read full blog post.

Pyroptosis: Mechanisms mediating cell death and pro-inflammatory cytokine release
By Victoria OsinskiPyroptosis is an inflammatory form of programmed cell death characterized by the release of pro-inflammatory cytokines IL-1 beta and IL-18.1,10 It is a process distinct from apoptosis and necrosis (T...  Read full blog post.

Caspase-3, The Executioner of Apoptosis
The Role of Caspase-3 in ApoptosisCaspase-3 enzyme is a member of the family of endoproteases which regulate inflammation and apoptosis signaling networks. Caspase-3 is known as an executioner caspase in apoptosis because of its role in coordinat...  Read full blog post.

VPS41 - An important regulator of lysosomal trafficking
Membrane fusion is an essential step during the trafficking of endosomes and vesicles throughout the cell. Membrane fusion events are facilitated by multisubunit tethering complexes (MTC) including CORVET and HOPS. These complexes interact with Rab...  Read full blog post.

Caspase-8 - a pro-apoptotic protein with dynamic roles in normal physiology and pathology
Caspases are a family of cysteine-aspartic acid proteases that are responsible for the initiation and execution of apoptosis. Caspase-8 is a 55 kDa protein expressed as an inactive procaspase that resides in the cytosol. Activation of Caspase-8 req...  Read full blog post.

Caspase 8 - a key mediator of apoptosis
Programmed cell death via apoptosis is a key controlled physiological process instigated by the cell death receptor family, their ligands, and the caspase cysteine protease family. All caspases exist in a precursor form that contains a prodomain, a...  Read full blog post.

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Bioinformatics

Gene Symbol CASP8