APAF-1 Overexpression Lysate

Images

 
Western Blot: APAF1 Overexpression Lysate (Adult Normal) [NBP2-04870] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for APAF1.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

APAF-1 Overexpression Lysate Summary

Description

APAF-1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC213433

Accession#: NM_181869

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
APAF1

Applications/Dilutions

Dilutions
  • Western Blot
Application Notes
This product is intended for use as a positive control in Western Blot.

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
37.8 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.This product is manufactured by and sold under license from OriGene Technologies and its use is limited solely for research purposes.

Alternate Names for APAF-1 Overexpression Lysate

  • APAF1
  • APAF-1
  • APAF-1apoptotic protease activating factor 1
  • apoptotic peptidase activating factor 1
  • apoptotic peptidase activating factor
  • apoptotic protease activating factor
  • apoptotic protease-activating factor 1
  • CED4
  • DKFZp781B1145
  • KIAA0413

Background

Apaf 1 (apoptosis protease-activating factor-1) is a key regulator of the mitochondrial apoptotic pathway (reviewed in Campioni et al, 2005), being the central element of the multimeric apoptosome. The apoptosome consists of cytochrome c, procaspase-9 and seven Apaf 1 monomers, and is considered to be core apoptotic machinery that executes mitochondria-dependent apoptosis. Cytochrome c, normally compartmentalized in the mitochorndria, is released into the cytoplasm following apoptotic stimuli. Apaf 1 binds cytochrome c in the cytoplasm and in the presence of dATP/ATP forms the apoptosome. The apoptosome binds procaspase-9 and promotes its autocatalytic activation. Active caspase-9, in turn, activates downstream caspases including 3, 6, and 7 contributing to the proteolytic caspase activation cascade which leads to cell death. Apaf 1 is an approx. 130 kDa protein. Alternative Apaf 1 splicing isoforms of Apaf 1 have been identified in mammalian cells, each having either or both an 11 amino acid insertion in the N-terminus and an additional WD40 repeat (43 aa) in the C-terminus; please refer to AceView of NCBI (http://www.ncbi.nih.gov/IEB/Research/Acembly/) for detailed information. Additionally, Apaf 1 can be proteolytically processed by active caspase-3 during apoptosis resulting in an Apaf 1 cleavage fragment of ~84 kDa (Lauber et al. 2001).

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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Product General Protocols

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Video Protocols

WB Video Protocol

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Research Areas for APAF-1 Lysate (NBP2-04870)

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Blogs on APAF-1.

Caspase 9 - an important apoptosis marker
Caspases are essential mediators of programmed cell death and are needed for both the induction of apoptosis as well as for aiding the degradation of cellular structures. Initiator caspases (such as Caspase-9) sense and respond to various signals i...  Read full blog post.

Heat Shock Proteins: An Overview
Heat Shock Proteins (HSPs) are a ubiquitous group of molecular chaperone proteins that have evolved unique mechanisms, within their host cells, to facilitate survival in hostile environments such as heat, oxidative (hypoxia), pH and cold. Under permis...  Read full blog post.

The Role of the Caspase 3 Antibody in Apoptosis Research
The caspases are a group of cysteine protease enzymes essential to apoptosis, inflammation and necrosis. Caspase 3 has been identified as one of the key mediators of apoptosis.Human caspases form three distinct groups: cytokine activators, which are...  Read full blog post.

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Bioinformatics

Gene Symbol APAF1