Overview

  • Product name
    Anti-ATPB antibody [4.3E8.D10]
    See all ATPB primary antibodies
  • Description
    Mouse monoclonal [4.3E8.D10] to ATPB
  • Host species
    Mouse
  • Specificity
    Detects the beta subunit of ATP synthase (ATPB) from mouse rat and human samples. This antibody is useful as a mitochondrial marker.
  • Tested applications
    Suitable for: WB, ICC, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Other Immunogen Type corresponding to ATPB. Intact rat mitochondria.

Properties

Applications

Our Abpromise guarantee covers the use of ab5432 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 - 2 µg/ml. Detects a band of approximately 57 kDa (predicted molecular weight: 57 kDa).
ICC 1/100 - 1/1000.
ICC/IF 1/100 - 1/1000.
IP Use a concentration of 2 - 5 µg/ml.

By immunoprecipatation, this antibody detects an 50 kDa protein representing ATP synthase (ATPB) from solubilized rat brain mitochondria.

Target

  • 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.
  • Sequence similarities
    Belongs to the ATPase alpha/beta chains family.
  • Cellular localization
    Mitochondrion. Mitochondrion inner membrane. Peripheral membrane protein.
  • Information by UniProt
  • Database links
  • Alternative names
    • ATP 5B antibody
    • ATP synthase H+ transporting mitochondrial F1 complex beta polypeptide antibody
    • ATP synthase subunit beta mitochondrial antibody
    • ATP synthase subunit beta, mitochondrial antibody
    • atp5b antibody
    • ATPB antibody
    • ATPB_HUMAN antibody
    • ATPMB antibody
    • ATPSB antibody
    • Epididymis secretory protein Li 271 antibody
    • HEL-S-271 antibody
    • Mitochondrial ATP synthase beta subunit antibody
    • Mitochondrial ATP Synthase Subunit Beta antibody
    • Mitochondrial ATP synthetase beta subunit antibody
    see all

Images

  • Immunocytochemistry/Immunofluorescent analysis of ATPB (red) in HEK293T cells. Cells fixed with 4% formaldehyde were permeabilized and blocked with 1X PBS containing 5% BSA and 0.3% Triton X-100 for 1 hour at room temperature. Cells were probed with ab5432 at 1:100 overnight at 4ºC in 1X PBS containing 1% BSA and 0.3% Triton X-100, washed with 1X PBS, and incubated with a fluorophore-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:200 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI. Images were taken on a Leica DM1000 microscope at 40X magnification.

  • Immunofluorescent analysis of ATPB in A431 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a ATPB monoclonal antibody (ab5432) at a dilution of 1:200 overnight at 4 C and incubated with a DyLight-488 conjugated secondary antibody. ATPB staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunofluorescent analysis of ATPB in HeLa cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a ATPB monoclonal antibody (ab5432) at a dilution of 1:200 overnight at 4 C and incubated with a DyLight-488 conjugated secondary antibody. ATPB staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

  • Immunoprecipatation of rat neuronal/glial cell extract using ab5432.
  • ICC/IF image of ab5432 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab5432, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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.

  • Immunocytochemistry/Immunofluorescence analysis of rat neuronal/glial cell culture using ab5432.

  • Anti-ATPB antibody [4.3E8.D10] (ab5432) at 1 µg/ml + Human testis tissue lysate - total protein (ab30257) at 20 µg

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 57 kDa
    Observed band size: 57 kDa


    Exposure time: 16 minutes

References

This product has been referenced in:
  • Diokmetzidou A  et al. Desmin and aB-crystallin interplay in the maintenance of mitochondrial homeostasis and cardiomyocyte survival. J Cell Sci 129:3705-3720 (2016). Read more (PubMed: 27566162) »
  • Laker RC  et al. The Mitochondrial Permeability Transition Pore Regulator Cyclophilin D Exhibits Tissue-Specific Control of Metabolic Homeostasis. PLoS One 11:e0167910 (2016). WB . Read more (PubMed: 28005946) »
See all 10 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer

Thank you for your email. Yes, we do carry human mitochondrial lysates as well: Human Liver Mitochondrial lysate (ab110339) Human Heart Mitochondrial lysate (ab110337) However, both lysates come in WB loading buffer (ready to be used in WB) and not iso-osmotic buffer as the bovine lysate. Regarding the antibodies: ab5432: This antibody has not been tested with bovine samples yet, but the sequence homology between rat (immunogen) and bovine is 96%. If you would like to try the antibody in bovine, you can take advantage of our testing discount program. For details please see below. ab110280: This antibody has been tested with bovine samples in WB and would be guaranteed for this species and application. Please see lane 2 for the WB data with bovine mitochondria. *Testing discount program: Here is some information about how the testing discount program works: The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount. *Abpromise guarantee: All our products are guaranteed for the tested species and applications as listed on the datasheet for 6 months after the purchase. For more details about our Abpromise guarantee please check this link: https://www.abcam.com/abpromise. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question
Answer

Merci pour votre appel de l'autre jour.

Je viens de recevoir la réponse du laboratoire. Malheureusement l'épitope du ab5432 n'as pas été déterminé non plus à notre connaissance.

Je suis désolée de ne pas pouvoir vous mieux aider en cette occasion. Veuillez ne pas hésiter à nous recontacter si vous avez des autres questions.

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Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Fruit fly (Drosophila melanogaster) Tissue lysate - whole (head)
Loading amount
30 µg
Specification
head
Gel Running Conditions
Reduced Denaturing
Blocking step
casein as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: 4°C

Ms. Zhiyin Xun

Verified customer

Submitted Aug 14 2007

Answer

Thank you for your enquiry and your patience. Based on the resources available for ATP Synthase Beta (ab5432), this product has not been epitope mapped yet. Occassionally customers will send us such information when they do the epitope mapping themselves, but I do not have any such information of this being done. Therefore, I would not know which domain was used as the immunogen. I apologize for not having this information.

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Answer

Thank you for your enquiry. The epitopes have not been determined for these two monoclonal antibodies. Ab5432 and ab14730 are different clones, and ab5432 was made using intact rat mitochondria as the immunogen, whereas human whole heart mitochondria was used as the immunogen for ab14730. Ab5432 was tested for application in IP and IF, and ab14730 was tested for application in Western blot and Immunocytochemistry. Please contact us again if you have any additional questions.

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Answer

Dr. Wang, Thank you for contacting us with your request. We do not have antibodies against all 13 of the proteins encoded by the mitochondrial genome yet. Right now we only seem to have an antibody against the COXI subunit, ab14705, which cross reacts with human, mouse, rat and cow. All the other antibodies we have against mitochondrial proteins are against ones encoded in the nucleus. I hope this is helpful to you, and keep checking with us as we add lots of new products each month and you may find what you need in the future.

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Answer

Thank you for your enquiry. We are very sorry to hear you are having problem with this product. We regret to inform you that ab5432has not been tested for Western blot application only for IP and IF. Please could you take a look at the on-line datasheet. It cross-reacts with human, mouse and rat protein ATP Synthase beta but we do not have any information if it recognizes S.cereviciae. We would suggest using rat brain lysate as positive control along with the samples. If you need anything further or any help then please let us know.

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Answer

We haven't tested this antibody in WB applications (ab5432 has been tested in Immunoprecipitation and Immunofluorescence), therefore we don't know. Although, by immunoprecipatation, this antibody detects an 50 kDa protein representing ATP synthase from solubilized rat brain mitochondria.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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