Our rapidly growing loading control range has taken a giant leap!
Due to huge demand, we are working on a new set of loading controls for different subcellular compartments. Our beta actin, tubulin and GAPDH loading control antibodies can be used for cytoplasmic and whole-cell extracts. However, for researchers working on specific organelle fractions, such as the nucleus and mitochondria, these are not suitable. Rather than making you search for a suitable marker protein and then for the relevant antibody which works well in western blots, we decided to do the hard work for you!
Lane 1 : Hela whole cell lysate
Lane 2 : Human skeletal muscle cell lysate
Lane 3 : Hela whole cell lysate + COX IV blocking peptide (ab16381)
Lane 4 : Human skeletal muscle cell lysate + COX IV blocking peptide (ab16381)
Rabbit polyclonal to COX IV (ab16056) 0.38 µg/ml
Alexa fluor Goat anti Rabbit IgG 1/10000
All lysates loaded at 20µg per well
COX IV blocking peptide (ab16381) used at 1µg/ml
Observed band size : 15kDa
The Voltage-Dependent Anion Channel (VDAC or mitochondrial porin) is an outer membrane mitochondrial protein. The VDAC protein is thought to form the major pores through which adenine nucleotides cross the outer mitochondrial membrane. VDAC has also been implicated in the formation of the mitochondrial permeability transition pore complex in apoptotic cells. This complex, formed by VDAC, adenine nucleotide translocator (ANT), and cyclophilin D (CypD), is thought to allow the mitochondria to undergo metabolic uncoupling and irreversible morphologic changes that ultimately destroy them during apoptosis.
Liu et al. Phospholipid scramblase 3 is the mitochondrial target of protein kinase C delta-induced apoptosis. Cancer Research 63: 1153–1156 (2003). PubMed: 14573790
Jonas et al. Proapoptotic N-truncated BCL-xL protein activates endogenous mitochondrial channels in living synaptic terminals. PNAS 101: 13590–13595 (2004). PubMed: 15342906
Cytochrome c oxidase, which is located in the inner mitochondrial membrane, is the terminal enzyme in the respiratory chain. COX functions as a dimer, with each monomer consisting of 13 subunits, 10 of which are nuclear encoded. COXIV is a nuclear-encoded subunit that may play a role in regulating COX activity.
COXIV is expressed at consistently high levels and thus makes a good loading control. However, be aware that many other proteins also run at 16 kDa. In these cases use VDAC1/Porin antibody (ab3434) instead.
ab14744 is described in:
Capaldi RA et al. Immunological approaches to the characterization and diagnosis of mitochondrial disease.
In press 2004. Mitochondrion
The ab14744 clone has been recently used in:
Kim TH et al. Bid-cardiolipin interaction at mitochondrial contact site contributes to mitochondrial cristae reorganization and cytochrome C release. Mol Biol Cell 15:3061-72 (2004). PubMed: 15107464
Hanson BJ et al. Cytochrome c oxidase-deficient patients have distinct subunit assembly profiles. J Biol Chem 276:16296-301 (2001). PubMed: 11278850
Soane L et al. C5b-9 terminal complement complex protects oligodendrocytes from death by regulating Bad through phosphatidylinositol 3-kinase/Akt pathway. J Immunol 167:2305-11 (2001). PubMed: 11490019
Liu M & Spremulli L Interaction of mammalian mitochondrial ribosomes with the inner membrane. J Biol Chem 275:29400-6 (2000). PubMed: 10887179
Atlante A et al. Cytochrome c is released from mitochondria in a reactive oxygen species (ROS)-dependent fashion and can operate as a ROS scavenger and as a respiratory substrate in cerebellar neurons undergoing excitotoxic death. J Biol Chem 275:37159-66 (2000). PubMed: 10980192
Liu H et al. Inhibition of c-Myc expression sensitizes hepatocytes to tumor necrosis factor-induced apoptosis and necrosis. J Biol Chem 275:40155-62 (2000). PubMed: 11016920
Zhou H et al. Akt regulates cell survival and apoptosis at a postmitochondrial level. J Cell Biol 151:483-94 (2000). PubMed: 11062251
Clark RS et al. Caspase-3 mediated neuronal death after traumatic brain injury in rats. J Neurochem 74:740-53 (2000). PubMed: 10646526
Wagenknecht B et al. Proteasome inhibitor-induced apoptosis of glioma cells involves the processing of multiple caspases and cytochrome c release. J Neurochem 75:2288-97 (2000). PubMed: 11080180