Immunofluorescent analysis of ATP Synthase Subunit beta was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ATP Synthase Subunit beta (3D5AB1) Mouse Monoclonal Antibody (A-21351) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bovine, C. elegans, Human, Mouse, Non-human primate, Rat|
|Published species reactivity||C. elegans, Fruit fly, Human, Mouse, Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Human Heart Mitochondria.|
|Storage buffer||HEPES buffered saline|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/mL|
|Immunofluorescence (IF)||2 µg/mL|
|Western Blot (WB)||0.5 µg/mL|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
ATP synthase is extremely conserved through evolution and can be found in plants, fungi, bacteria, and animals. The ATP synthase enzyme is a transmembrane protein responsible for driving the reversible reaction from ADP+ phosphate to ATP. This reaction is accomplished by a flux of protons across the membrane as a result of electron transfer.
The ATP synthase protein has two main sections; the F1 ATP-ase (soluble) and the F0 ATP-ase (membrane embedded). The F1 section consists of the alpha, beta, gamma, delta, and epsilon subunits. While the F0 consists of a, b, and c subunits.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Assembly of human mitochondrial ATP synthase through two separate intermediates, F1-c-ring and b-e-g complex.
A-21351 was used in western blot to study the assembly process of human mitochondrial ATP synthase
|Fujikawa M,Sugawara K,Tanabe T,Yoshida M||FEBS letters (589:2707)||2015|
Quantitative analysis of PPT1 interactome in human neuroblastoma cells.
A-21351 was used in western blot to examine palmitoyl protein thioesterase.
|Scifo E,Szwajda A,Soliymani R,Pezzini F,Bianchi M,Dapkunas A,D¿bski J,Uusi-Rauva K,Dadlez M,Gingras AC,Tyynelä J,Simonati A,Jalanko A,Baumann MH,Lalowski M||Data in brief (4:207)||2015|
Proteomic analysis of the palmitoyl protein thioesterase 1 interactome in SH-SY5Y human neuroblastoma cells.
A-21351 was used in western blot to identify substrates of human PPT using neuronal cells.
|Scifo E,Szwajda A,Soliymani R,Pezzini F,Bianchi M,Dapkunas A,D¿bski J,Uusi-Rauva K,Dadlez M,Gingras AC,Tyynelä J,Simonati A,Jalanko A,Baumann MH,Lalowski M||Journal of proteomics (123:42)||2015|
|Fruit fly||Not Cited||
Involvement of ATP synthase ß subunit in chikungunya virus entry into insect cells.
A-21351 was used in western blot to identify the ATP synthase beta subunit as a mosquito-cell-expressed chikungunya virus-binding protein
|Fongsaran C,Jirakanwisal K,Kuadkitkan A,Wikan N,Wintachai P,Thepparit C,Ubol S,Phaonakrop N,Roytrakul S,Smith DR||Archives of virology (159:3353)||2014|
High-fat diet-induced impairment of skeletal muscle insulin sensitivity is not prevented by SIRT1 overexpression.
A-21351 was used in western blot to test if constitutive activation of SIRT1 in skeletal muscle prevents high fat diet-induced muscle insulin resistance
|White AT,Philp A,Fridolfsson HN,Schilling JM,Murphy AN,Hamilton DL,McCurdy CE,Patel HH,Schenk S||American journal of physiology. Endocrinology and metabolism (307:E764)||2014|
A Drosophila model of mitochondrial disease caused by a complex I mutation that uncouples proton pumping from electron transfer.
A-21351 was used in western blot to elucidate the mechanisms by which complex I dysfunction contribute to mitochondrial disease
|Burman JL,Itsara LS,Kayser EB,Suthammarak W,Wang AM,Kaeberlein M,Sedensky MM,Morgan PG,Pallanck LJ||Disease models and mechanisms (7:1165)||2014|
IF1, a natural inhibitor of mitochondrial ATP synthase, is not essential for the normal growth and breeding of mice.
A-21351 was used in western blot to characterize a mouse strain in which IF gene was destroyed.
|Nakamura J,Fujikawa M,Yoshida M||Bioscience reports (33:null)||2013|
Advanced oxidation protein products induce cardiomyocyte death via Nox2/Rac1/superoxide-dependent TRAF3IP2/JNK signaling.
A-21351 was used in western blot to identify the mechanism for the cardiomyocyte death induced by advanced oxidation protein products.
|Valente AJ,Yoshida T,Clark RA,Delafontaine P,Siebenlist U,Chandrasekar B||Free radical biology and medicine (60:125)||2013|
Inhibition of tumor cell surface ATP synthesis by pigment epithelium-derived factor: implications for antitumor activity.
A-21351 was used in western blot to study the interactions between the extracellular PEDF and tumor cell surface ATP synthase.
|Deshpande M,Notari L,Subramanian P,Notario V,Becerra SP||International journal of oncology (41:219)||2012|
Assessing actual contribution of IF1, inhibitor of mitochondrial FoF1, to ATP homeostasis, cell growth, mitochondrial morphology, and cell viability.
A-21351 was used in western blot to study energy metabolism in IF knockdown cells.
|Fujikawa M,Imamura H,Nakamura J,Yoshida M||The Journal of biological chemistry (287:18781)||2012|
Examining the interactome of huperzine A by magnetic biopanning.
A-21351 was used in western blot to examine the interactome of Huperzine A.
|Guo W,Liu S,Peng J,Wei X,Sun Y,Qiu Y,Gao G,Wang P,Xu Y||PloS one (7:null)||2012|
|Not Applicable||Not Cited||
Modes of metabolic compensation during mitochondrial disease using the Drosophila model of ATP6 dysfunction.
A-21351 was used in western blot to investigate metabolic changes in a Drosophila model of human mitochondrial encephalomyopathy
|Celotto AM,Chiu WK,Van Voorhies W,Palladino MJ||PloS one (6:null)||2011|
The PGC-1¿-related coactivator promotes mitochondrial and myogenic adaptations in C2C12 myotubes.
A-21351 was used in western blot to examine the effect of PGC-α-related coactivator on the metabolic profile of C2C2 myotubes.
|Philp A,Belew MY,Evans A,Pham D,Sivia I,Chen A,Schenk S,Baar K||American journal of physiology. Regulatory, integrative and comparative physiology (301:R864)||2011|
Knockdown of DAPIT (diabetes-associated protein in insulin-sensitive tissue) results in loss of ATP synthase in mitochondria.
A-21351 was used in western blot to investigate the contribution of diabetes-associated protein in insulin-sensitive tissue to ATP synthesis.
|Ohsakaya S,Fujikawa M,Hisabori T,Yoshida M||The Journal of biological chemistry (286:20292)||2011|
The microbiome and butyrate regulate energy metabolism and autophagy in the mammalian colon.
A-21351 was used in western blot to study the effects of microbiome-derived butyrate on the energy metabolism of colonocytes.
|Donohoe DR,Garge N,Zhang X,Sun W,O'Connell TM,Bunger MK,Bultman SJ||Cell metabolism (13:517)||2011|
The proteome of human brain after ischemic stroke.
A-21351 was used in western blot to determine the proteome of the ischemic human brain.
|Cuadrado E,Rosell A,Colomé N,Hernández-Guillamon M,García-Berrocoso T,Ribo M,Alcazar A,Ortega-Aznar A,Salinas M,Canals F,Montaner J||Journal of neuropathology and experimental neurology (69:1105)||2010|
|Not Applicable||Not Cited||
WNT1-inducible signaling pathway protein-1 activates diverse cell survival pathways and blocks doxorubicin-induced cardiomyocyte death.
A-21351 was used in western blot to identify doxorubicin-induced alterations in adult mouse cardiomyocytes
|Venkatesan B,Prabhu SD,Venkatachalam K,Mummidi S,Valente AJ,Clark RA,Delafontaine P,Chandrasekar B||Cellular signalling (22:809)||2010|
|Not Applicable||Not Cited||
Bz-423 superoxide signals B cell apoptosis via Mcl-1, Bak, and Bax.
A-21351 was used in western blot to describe the signaling pathway engaged by Bz-423 in a Burkitt lymphoma cell line
|Blatt NB,Boitano AE,Lyssiotis CA,Opipari AW,Glick GD||Biochemical pharmacology (78:966)||2009|
|Not Applicable||Not Cited||
Bz-423 superoxide signals apoptosis via selective activation of JNK, Bak, and Bax.
A-21351 was used in western blot to identify the signaling pathway activated by Bz-423 in mouse embryonic fibroblasts
|Blatt NB,Boitano AE,Lyssiotis CA,Opipari AW,Glick GD||Free radical biology and medicine (45:1232)||2008|
Mitochondrial proteomics of the retinal pigment epithelium at progressive stages of age-related macular degeneration.
A-21351 was used in western blot to examine the subproteome of mitochondria isolated from human donor retinal pigment epithelium graded with the Minnesota Grading System
|Nordgaard CL,Karunadharma PP,Feng X,Olsen TW,Ferrington DA||Investigative ophthalmology and visual science (49:2848)||2008|
Motor neuron degeneration in amyotrophic lateral sclerosis mutant superoxide dismutase-1 transgenic mice: mechanisms of mitochondriopathy and cell death.
A-21351 was used in western blot to examine the neuronal cell death in the spinal cords of mSOD1 and wtSOD1 mice.
|Martin LJ,Liu Z,Chen K,Price AC,Pan Y,Swaby JA,Golden WC||The Journal of comparative neurology (500:20)||2007|
|Not Applicable||Not Cited||
Coordination of nuclear- and mitochondrial-DNA encoded proteins in cancer and normal colon tissues.
A-21351 was used in western blot to investigate how oxidative phosphorylation contributes to the coordination between nuclear- and mitochondria-encoded mitochondrial proteins
|Mazzanti R,Giulivi C||Biochimica et biophysica acta (1757:618)||2006|
In vivo time-lapse imaging of mitochondria in healthy and diseased peripheral myelin sheath.
A-21351 was used in immunohistochemistry to study mitochondrial dynamics and morphology in myelinating Schwann cells in living mice.
|Gonzalez S,Fernando R,Berthelot J,Perrin-Tricaud C,Sarzi E,Chrast R,Lenaers G,Tricaud N||Mitochondrion (23:32)||2015|
Apolipoprotein A-I enhances proliferation of human endothelial progenitor cells and promotes angiogenesis through the cell surface ATP synthase.
A-21351 was used in flow cytometry and immunocytochemistry to examine the effect of apoA-I in the proliferation and the angiogenic capacity of early hEPC.
|González-Pecchi V,Valdés S,Pons V,Honorato P,Martinez LO,Lamperti L,Aguayo C,Radojkovic C||Microvascular research (98:9)||2015|
PINK1-Parkin pathway activity is regulated by degradation of PINK1 in the mitochondrial matrix.
A-21351 was used in western blot to investigate how PINK1 is degraded.
|Thomas RE,Andrews LA,Burman JL,Lin WY,Pallanck LJ||PLoS genetics (10:null)||2014|
The N-terminal helix of Bcl-xL targets mitochondria.
A-21351 was used in western blot to test if the N-terminus of Bcl-xL contains a mitochondrial targeting signal.
|McNally MA,Soane L,Roelofs BA,Hartman AL,Hardwick JM||Mitochondrion (13:119)||2013|
Identification of human fumarylacetoacetate hydrolase domain-containing protein 1 (FAHD1) as a novel mitochondrial acylpyruvase.
A-21351 was used in western blot to determine the enzymatic function of FAHD1.
|Pircher H,Straganz GD,Ehehalt D,Morrow G,Tanguay RM,Jansen-Dürr P||The Journal of biological chemistry (286:36500)||2011|
The GTPase RAB20 is a HIF target with mitochondrial localization mediating apoptosis in hypoxia.
A-21351 was used in western blot to characterize Rab20, a hypoxia-induced gene
|Hackenbeck T,Huber R,Schietke R,Knaup KX,Monti J,Wu X,Klanke B,Frey B,Gaipl U,Wullich B,Ferbus D,Goubin G,Warnecke C,Eckardt KU,Wiesener MS||Biochimica et biophysica acta (1813:1)||2011|
A sensitive, simple assay of mitochondrial ATP synthesis of cultured mammalian cells suitable for high-throughput analysis.
A-21351 was used in western blot to establish an assay for measuring mitochondrial ATP synthesis in cultured mammalian cells
|Fujikawa M,Yoshida M||Biochemical and biophysical research communications (401:538)||2010|
Pyruvate suppresses PGC1alpha expression and substrate utilization despite increased respiratory chain content in C2C12 myotubes.
A-21351 was used in western blot to study the effects of osmotic changes on the regulation of mitochondrial metabolism and biogenesis in C2C12 myotubes
|Philp A,Perez-Schindler J,Green C,Hamilton DL,Baar K||American journal of physiology. Cell physiology (299:C240)||2010|
Population of ATP synthase molecules in mitochondria is limited by available 6.8-kDa proteolipid protein (MLQ).
A-21351 was used in immunocytochemistry to investigate MLQ as a regulator of mitochondrial ATP synthesis.
|Fujikawa M,Ohsakaya S,Sugawara K,Yoshida M||Genes to cells : devoted to molecular and cellular mechanisms (19:153)||2014|
|Not Applicable||2 µg/ml||
Histochemical methods for the diagnosis of mitochondrial diseases.
A-21351 was used in immunocytochemistry, immunohistochemistry - frozen section, and immunohistochemistry - paraffin section to review methods to study oxidative phosphorylation
|De Paepe B,De Bleecker JL,Van Coster R||Current protocols in human genetics (Chapter 19:null)||2009|
|Not Applicable||0.5-4 µg/ml||
Subdiffraction-resolution fluorescence imaging of proteins in the mitochondrial inner membrane with photoswitchable fluorophores.
A-21351 was used in immunocytochemistry to show that commercially available standard fluorescent probes are efficient photoswitches
|van de Linde S,Sauer M,Heilemann M||Journal of structural biology (164:250)||2008|
|Not Applicable||Not Cited||
2,2'-thiodiethanol: a new water soluble mounting medium for high resolution optical microscopy.
A-21351 was used in immunocytochemistry to characterize a new water soluble mounting medium called 2,2'-thiodiethanol, that is used for high resolution optical microscopy
|Staudt T,Lang MC,Medda R,Engelhardt J,Hell SW||Microscopy research and technique (70:1)||2007|
Heterogeneous levels of oxidative phosphorylation enzymes in rat adrenal glands.
A-21351 was used in immunohistochemistry - paraffin section to observe and characterize mitochondria in the rat adrenal gland
|Ogawa K,Harada K,Endo Y,Sagawa S,Inoue M||Acta histochemica (113:24)||2011|