Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
Mouse monoclonal ATPB antibody [3D5]. Validated in WB, IP, IHC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Cow, Human, Pig, Caenorhabditis elegans, Monkey. Cited in 131 publication(s).
- Datasheet
- References (142)
- Protocols
Overview
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Product nameAnti-ATPB antibody [3D5] - Mitochondrial Marker
See all ATPB primary antibodies -
DescriptionMouse monoclonal [3D5] to ATPB - Mitochondrial Marker
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Host speciesMouse
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SpecificityHuman and Bovine complex V beta subunit (ATPB).
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Tested applicationsSuitable for: WB, IHC-Fr, ICC/IF, IP, Flow Cyt, IHC-Pmore details
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Species reactivityReacts with: Mouse, Rat, Cow, Human, Pig, Caenorhabditis elegans, Monkey
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Immunogen
Whole heart mitochondria (Human).
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General notes
This antibody clone is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
Properties
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FormLiquid
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Storage instructionsShipped 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.
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Storage bufferPreservative: 0.02% Sodium azide
Constituent: HEPES buffered saline -
Concentration information loading... -
PurityIgG fraction
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Purification notesNear homogeneity as judged by SDS-PAGE. The antibody was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation.
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ClonalityMonoclonal
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Clone number3D5
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IsotypeIgG1
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Light chain typekappa
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Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
Our Abpromise guarantee covers the use of ab14730 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 0.5 µg/ml. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa). | |
| IHC-Fr | 1/100. | |
| ICC/IF | Use a concentration of 1 - 2 µg/ml. | |
| IP | Use at an assay dependent concentration. PubMed: 21490313 | |
| Flow Cyt | Use a concentration of 1 µg/ml. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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| IHC-P | Use at an assay dependent concentration. |
Target
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FunctionMitochondrial 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.
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Sequence similaritiesBelongs to the ATPase alpha/beta chains family.
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Cellular localizationMitochondrion. Mitochondrion inner membrane. Peripheral membrane protein.
- Information by UniProt
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Database links
- Entrez Gene: 327675 Cow
- Entrez Gene: 506 Human
- Entrez Gene: 11947 Mouse
- Entrez Gene: 100157156 Pig
- Entrez Gene: 171374 Rat
- GenBank: BC016512 Human
- Omim: 102910 Human
- SwissProt: P00829 Cow
see all -
Alternative names
- ATP 5B antibody
- ATP synthase H+ transporting mitochondrial F1 complex beta polypeptide antibody
- ATP synthase subunit beta mitochondrial antibody
see all
Images
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![Western blot - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)](https://www.abcam.com/ps/products/14/ab14730/Images/ab14730-323620-anti-atpb-antibody-3d5-mitochondrial-marker-western-blot.jpg)
Western blot - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)All lanes :
Lane 1 : HeLa
Lane 2 : HepG2
Lane 3 : Hu liver
Lane 4 : Ms liver
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 52 kDaMerged signal (red and green). Green - ab14730 observed at 52 kDa. Red - loading control, ab181602 observed at 37 kDa.
Samples were subjected to SDS-PAGE. ab14730 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1ug/mL and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)](https://www.abcam.com/ps/products/14/ab14730/Images/ab14730-22076-anti-atpb-antibody-3d5-mitochondrial-marker-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)ab14730 staining ATPB in cultured human fibroblasts. Cells were fixed, permeabilized and then labelled with ab14730 followed by an AlexaFluor® 488-conjugated Goat anti-Mouse IgG1-specific secondary antibody (2 µg/ml) -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)](https://www.abcam.com/ps/products/14/ab14730/Images/ab14730-22078-anti-atpb-antibody-3d5-mitochondrial-marker-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)ab14730 (2µg/ml) staining ATPB in human duodenum using an automated system (DAKO Autostainer Plus). Using this protocol there is cytoplasmic and mitochondrial staining of epithelium.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required. -
![Flow Cytometry - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)](https://www.abcam.com/ps/products/14/ab14730/Images/ab14730-22075-anti-atpb-antibody-3d5-mitochondrial-marker-flow-cytometry.jpg)
Flow Cytometry - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)ab14730 (blue) at 1µg/ml staining ATPB in HL-60 cells and analyzed by Flow cytometry. Red histogram represents equal quantity of isotype control. -
![Western blot - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)](https://www.abcam.com/ps/products/14/ab14730/Images/ab14730-22077-anti-atpb-antibody-3d5-mitochondrial-marker-western-blot.jpg)
Western blot - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)All lanes : Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)
Lane 1 : isolated mitochondria from human heart at 5 µg
Lane 2 : isolated mitochondria from bovine heart at 1 µg
Lane 3 : isolated mitochondria from rat heart at 10 µg
Lane 4 : isolated mitochondria from mouse heart at 10 µg
Predicted band size: 52 kDa
Observed band size: 52 kDa -
![Immunohistochemistry (Frozen sections) - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)](https://www.abcam.com/ps/products/14/ab14730/Images/ab14730-230546-anti-atpb-antibody-3d5-mitochondrial-marker-immunohistochemistry.jpg)
Immunohistochemistry (Frozen sections) - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)This image is courtesy of an anonymous Abreviewab14730 staining ATPB in rat liver tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/100 in 10% goat serum) for 18 hours at 4°C. An Alexa Fluor® 555-conjugated goat anti-mouse polyclonal (1/500) was used as the secondary antibody.
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![Western blot - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)](https://www.abcam.com/ps/products/14/ab14730/Images/ab14730-22079-anti-atpb-antibody-3d5-mitochondrial-marker-western-blot.jpg)
Western blot - Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730)All lanes : Anti-ATPB antibody [3D5] - Mitochondrial Marker (ab14730) at 0.8 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : Human liver tissue lysate - total protein (ab29889)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Predicted band size: 52 kDa
Observed band size: 52 kDa
Protocols
References
This product has been referenced in:
- Cummins N et al. Disease-associated tau impairs mitophagy by inhibiting Parkin translocation to mitochondria. EMBO J 38:N/A (2019). Read more (PubMed: 30538104) »
- Peel S et al. Introducing an automated high content confocal imaging approach for Organs-on-Chips. Lab Chip 19:410-421 (2019). Read more (PubMed: 30663729) »
