Anti-ATP5A antibody (ab151229)
Key features and details
- Rabbit polyclonal to ATP5A
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-ATP5A antibody
See all ATP5A primary antibodies -
Description
Rabbit polyclonal to ATP5A -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Chicken, Cow, Chimpanzee, Orangutan
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Immunogen
Synthetic peptide corresponding to Human ATP5A aa 200-300 conjugated to keyhole limpet haemocyanin.
(Peptide available asab151534) -
Positive control
- WB: Human fetal brain and human fetal heart tissue lysates and Raw264.7, HEK293, MCF7, HepG2, HL60 and PC12 whole cell lysates. IHC-P: Human heart muscle tissue sections. ICC/IF: HeLa cells.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab151229 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application Abreviews Notes WB Use a concentration of 1 µg/ml. Detects a band of approximately 59 kDa (predicted molecular weight: 59 kDa). IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. ICC/IF Use a concentration of 1 µg/ml. Target
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Function
Mitochondrial membrane ATP synthase (F(1)F(0) ATP synthase or Complex V) produces ATP from ADP in the presence of a proton gradient across the membrane which is generated by electron transport complexes of the respiratory chain. F-type ATPases consist of two structural domains, F(1) - containing the extramembraneous catalytic core, and F(0) - containing the membrane proton channel, linked together by a central stalk and a peripheral stalk. During catalysis, ATP synthesis in the catalytic domain of F(1) is coupled via a rotary mechanism of the central stalk subunits to proton translocation. Subunits alpha and beta form the catalytic core in F(1). Rotation of the central stalk against the surrounding alpha(3)beta(3) subunits leads to hydrolysis of ATP in three separate catalytic sites on the beta subunits. Subunit alpha does not bear the catalytic high-affinity ATP-binding sites. -
Tissue specificity
Fetal lung, heart, liver, gut and kidney. Expressed at higher levels in the fetal brain, retina and spinal cord. -
Sequence similarities
Belongs to the ATPase alpha/beta chains family. -
Post-translational
modificationsThe N-terminus is blocked. -
Cellular localization
Mitochondrion inner membrane. Peripheral membrane protein. - Information by UniProt
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Database links
- Entrez Gene: 455397 Chimpanzee
- Entrez Gene: 282578 Cow
- Entrez Gene: 498 Human
- Entrez Gene: 11946 Mouse
- Entrez Gene: 100173854 Orangutan
- Entrez Gene: 65262 Rat
- Omim: 164360 Human
- SwissProt: A5A6H5 Chimpanzee
see all -
Alternative names
- ATP synthase alpha chain antibody
- ATP synthase alpha chain, mitochondrial antibody
- ATP synthase subunit alpha antibody
see all
Images
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Western blot - Anti-ATP5A antibody (ab151229)All lanes : Anti-ATP5A antibody (ab151229) at 1 µg/ml
Lane 1 : Brain (Human) Tissue Lysate - fetal normal tissue (ab29467)
Lane 2 : Heart (Human) Whole Cell Lysate - fetal normal tissue
Lane 3 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate
Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 6 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 7 : HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate
Lane 8 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 59 kDa
Observed band size: 59 kDa
Additional bands at: 37 kDa, 50 kDa, 75 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 10 seconds
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab151229 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution. -
Immunocytochemistry/ Immunofluorescence - Anti-ATP5A antibody (ab151229)ICC/IF image of ab151229 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab151229, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
This antibody also gave a positive result in 100% methanol fixed (5 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 4% formaldehyde fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 1µg/ml.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP5A antibody (ab151229)IHC image of ATP5A staining in human heart muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab151229, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Protocols
Datasheets and documents
References (4)
ab151229 has been referenced in 4 publications.
- Piro-Mégy C et al. Dominant mutations in mtDNA maintenance gene SSBP1 cause optic atrophy and foveopathy. J Clin Invest 130:143-156 (2020). PubMed: 31550237
- Pajak A et al. Defects of mitochondrial RNA turnover lead to the accumulation of double-stranded RNA in vivo. PLoS Genet 15:e1008240 (2019). PubMed: 31365523
- Arena G et al. Mitochondrial MDM2 Regulates Respiratory Complex I Activity Independently of p53. Mol Cell 69:594-609.e8 (2018). PubMed: 29452639
- Collins CM et al. ATP synthase F1 subunits recruited to centromeres by CENP-A are required for male meiosis. Nat Commun 9:2702 (2018). PubMed: 30006572
Images
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Western blot - Anti-ATP5A antibody (ab151229)All lanes : Anti-ATP5A antibody (ab151229) at 1 µg/ml
Lane 1 : Brain (Human) Tissue Lysate - fetal normal tissue (ab29467)
Lane 2 : Heart (Human) Whole Cell Lysate - fetal normal tissue
Lane 3 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate
Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 6 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 7 : HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate
Lane 8 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 59 kDa
Observed band size: 59 kDa
Additional bands at: 37 kDa, 50 kDa, 75 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 10 seconds
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab151229 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution. -
Immunocytochemistry/ Immunofluorescence - Anti-ATP5A antibody (ab151229)
ICC/IF image of ab151229 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab151229, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
This antibody also gave a positive result in 100% methanol fixed (5 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 4% formaldehyde fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 1µg/ml.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP5A antibody (ab151229)IHC image of ATP5A staining in human heart muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab151229, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
