459240

antibody from Invitrogen Antibodies

Targeting: ATP5F1A ATP5A, ATP5A1, ATP5AL2, ATPM, hATP1, OMR, ORM

Provider product page for 459240

Western blot

Immunocytochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [37]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [1]
Other assay [17]

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Product number: 459240 - Provider product page
Provider: Invitrogen Antibodies
Product name: ATP5A1 Monoclonal Antibody (7H10BD4F9)
Antibody type: Monoclonal
Antigen: Other
Reactivity: Human, Mouse, Rat, Bovine
Host: Mouse
Isotype: IgG
Antibody clone number: 7H10BD4F9
Vial size: 100 µL
Concentration: 1.0 mg/mL
Storage: 4° C

ATP5F1A protein structure - 459240 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of ATP5A was achieved by transfecting Hep G2 with ATP5A specific siRNAs (Silencer® select Product # s1769). Western blot analysis (Fig. a) was performed using membrane enriched extracts from the ATP5A knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with ATP5A1 Monoclonal Antibody (7H10BD4F9) (Product # 459240, 1µg/mL and Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to ATP5A.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-ATP5A Mouse Monoclonal Antibody (Product # 459240) and a 54 kDa band corresponding to an isoform of ATP5A was observed in cell lines and tissues tested. Membrane enriched extracts (30 µg lysate) of Hep G2 (Lane 1), HeLa (Lane 2), Caco-2 (Lane 3), HCT 116 (Lane 4), Mouse Heart (Lane 5), Rat Heart (Lane 6), Mouse Thymus (Lane 7) and Mouse Lung (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A 25 kDa band (*) corresponding to circulting tissue IgG was observed in mouse tissues.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 F1FO-ATP synthase deregulation in 5xFAD mouse synaptic mitochondrial. ( a ) Synaptic mitochondria from 5xFAD mice demonstrated an age-dependent decrease in their respiratory control ratio (RCR). Six nonTg and 5 5xFAD mice at 4 months old and 6 nonTg and 5 5xFAD mice at 9 months old were used. ( b ) Synaptic mitochondria from 5xFAD mice had an age-dependent decrease in ATP synthesis. Six nonTg and 6 5xFAD mice at 4 months old and those from six nonTg and seven 5xFAD mice at 9 months old were used in the experiments. ( c ) Synaptic mitochondria from 5xFAD mice demonstrated an early decrease in the F1FO-ATP synthase catalytic activity at 4 months old which was exacerbated at 9 months old. Five mice of each group at 4 months old and seven nonTg and nine 5xFAD mice at 9 months old were used in the experiment. ( d ) Age-dependent increase in oligomycin-insensitive respiration of synaptic mitochondria from 5xFAD mice. Six nonTg and five 5xFAD mice at 4 months old, and six nonTg and five 5xFAD mice at 9 months old were used in the experiments. ( e , f ) Decreased oligomycin sensitivity of synaptic mitochondria from 5xFAD mice at 4 ( e ) and 9 months old ( f ). All data are presented as percentage of the activity of the corresponding vehicle-treated mitochondrial fractions. Six nonTg and five 5xFAD mice at 4 months old, and seven nonTg and seven 5xFAD mice at 9 months old were used in the experiments. ( g , h ) Increased F1 dissociation in synaptic mitochondria from 5xFAD mic

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 OSCP overexpression ameliorates Abeta-induced mitochondrial dysfunction in mouse neurons. Mouse cortical neurons were exposed to 500 nM oligomeric Abeta1-42 for 24 h. ( a ) Attenuated oligomeric Abeta1-42-induced OSCP reduction by OSCP overexpression. Western blot images show OSCP expression level in mouse primary neurons which is representative of seven samples in each group. ( b ) Shows immunofluorescent staining of OSCP (green) in primary cultured neurons. COXIV (red) was used to determine the localization of OSCP in mitochondria. * P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 8 OSCP overexpression ameliorates Abeta-induced mitochondrial dysfunction in human neurons. ( a ) Human neurons were exposed to 500 nM oligomeric Abeta1-42 for 4 days, then subjected to immunoblotting detection of OSCP levels. Abeta induced a significant reduction in OSCP expression level. The lower panel shows representative immunoblotting images. Tom40 was used as the loading control. n =4-5 samples of each group. ( b ) Co-immunoprecipitation of OSCP and Abeta in Abeta-treated human neurons. ( c ) Significantly increased OSCP expression in the OSCP overexpressing neurons. The lower panel shows representative immunoblotting images. Tom40 was used as the loading control. n =4 samples of each group. ( d ) Preserved ATP production by OSCP overexpression. * P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 9 PARP-1 protein levels in mitochondrial and nuclear fractions of myoblasts and myotubes. Western blot analysis detected elevated expression of PARP-1 in myoblasts than in myotubes for both nuclear (A) and mitochondrial (B) fractions. Densitometric analysis of PARP-1 protein level in myoblast was set as 100%. Statistical analyses of n = 3 independent experiments were assessed; where ** indicates p

Supportive validation

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Experimental details: Fig. 2. Assessment of mitochondrial bioenergetics in neuronal tissues (brainstem, posterior columns, nerve roots and dorsal root ganglia). A quantitative western blot assay was developed to measure FXN (A), COXI and COXII (B), cytochrome c (C), ATP synthase (D) and TOM22 (E) in neuronal tissues [brainstem, posterior columns (PC), nerve roots and dorsal root ganglia (DRG)]. (A) Representative western blot of human FXN expression in YG8R mice in the four neuronal tissues used in the study. Human FXN was only detected in YG8R mice, in which the transgene was present, and not in wild type (WT; data not shown). Western blot results were quantified for each lane using Fujifilm's Multi-Gauge software. To allow for loading variation, values were normalized to the actin control. Final values were expressed as a ratio to the value of FXN expression in nerve roots. DRG and nerve roots (distal tissues) expressed less FXN than the posterior columns and brainstem (proximal tissues). Values are expressed as mean+-s.e.m.; *** P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 Expression of the mitochondrial marker ATP synthase is higher in gastrocnemius muscle in PNA rats. Gastrocnemius ( A - C ), tibialis anterior ( D - F ), and soleus ( G - I ) skeletal muscles from 16-week-old female rats exposed to sesame oil ( n = 4-5) or testosterone ( n = 4-5) were cryosectioned at 20 um thickness prior to indirect immunolabeling for ATP synthase. Scale bars, 200 um. For each rat, 3 distinct fields per muscle sample were imaged and the fluorescence intensity quantified ( C , F , I ). Fluorescence intensity data were analyzed using a Mann-Whitney U test. Fluorescence intensity trended higher in muscles from PNA rats for gastrocnemius (# p = 0.0556), trended slightly lower in TA (# p = 0.0979), but was not significantly different in soleus muscle. Quartiles with the median are shown. ( J - L ) Protein expression was examined by Western blot and quantified by densitometric analysis. Samples of ( J ) GN, ( K ) TA, and ( L ) SOL muscles from rats exposed to sesame oil ( n = 8) or testosterone ( n = 8) were separated by SDS-PAGE and transferred to nitrocellulose prior to detection with antibodies to the alpha subunit of ATP synthase or GAPDH (top panels). Values for ATP synthase were normalized to GAPDH and analyzed using an unpaired t test or Mann-Whitney U test, which indicated significantly greater expression of ATP synthase in GN, but not TA or SOL, from PNA rats (bottom panels). Values shown are means with SD. ** p < 0.005.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 4 Citrate synthase and complex I activities and expression levels of single subunit proteins of respiratory complexes I-V in R349P desmin knock-in mice. a Determination of the citrate synthase activity by a spectrophotometric assay using identical amounts of total protein extracts. Note the marked activity reduction in homozygous mice. For this approach, soleus muscles obtained from five mice of each genotype (WT, HET, HOM) were pooled and subjected to four-time repeated measurements. Column chart shows mean values of these technical replicates. b Determination of complex I activity by a colorimetric assay normalized to muscle weight. Though this analysis, performed in duplicate on non-pooled samples derived from three animals per genotype, showed a reduction of complex I activity in homozygous mice, the data failed to reach statistical significance. An additional normalization of the obtained complex I values to the citrate synthase values resulted in similar activity levels for the three genotypes (data not shown). Column chart shows mean values and standard errors. c Immunoblotting using antibodies directed against specific complex I-V proteins revealed increased, decreased, and missing signals of complex I subunit proteins in heterozygous and homozygous mice, whereas the signal intensities of complex II-V subunit proteins showed no obvious differences. Desmin immunoblotting confirmed a markedly decreased level of mutant desmin in homozygous mice. In heteroz

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Pcp1p mutants have altered steady-state levels for select adenosine triphosphate (ATP) synthase subunits . (A) Nonfunctional pcp1 alleles have reduced levels of ATP synthase alpha subunit. Proteins from isolated mitochondria were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunoblot analysis using anti-ATPV alpha subunit and anti-Tom40 antibodies. Tom40p was used as a control to confirm equal loading of samples. (B) Densitometric analysis of the levels of ATP synthase alpha subunit relative to Tom40p and normalized to the PCP1 wild-type control. Data presented as the average +- standard deviation from four experiments. * P = 0.03 for Delta pcp1 and 0.05 for G315D). (C) Tim11p levels vary across the pcp1 mutants. Proteins isolated as described above were analyzed by immunoblot for Tim11p and anti-Tom40 levels. (D) Densitometric analysis of the level of Tim11p relative to Tom40p and normalized to the PCP1 wild-type strain. Data represent the average +- standard deviation from six experiments. * P = 0.008.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 Nonfunctional pcp1 alleles have reduced adenosine triphosphate (ATP) synthase super-complexes formed . ATP synthase complex oligomerization and ATPase activity were measured following blue native-polyacrylamide gel electrophoresis (PAGE). (A) Representative immunoblot showing ATP synthase oligomerization status using ATP synthase alpha subunit antibodies. Mitochondrial proteins from the strains indicated were solubilized in buffer at a digitonin to protein ratio of 1.4:1 (g/g) and 0.7:1 (g/g). (B) The corresponding sodium dodecyl sulfate PAGE (SDS-PAGE) of Tom40p levels by immunoblot analysis for samples presented in (A) confirm equal loading. (C) Representative in-gel ATP synthase activity for the same pcp1 strains under the two solubilization conditions described above ( n = 3).