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).

Overview

  • Product name
    Anti-ATPB antibody [3D5] - Mitochondrial Marker
    See all ATPB primary antibodies
  • Description
    Mouse monoclonal [3D5] to ATPB - Mitochondrial Marker
  • Host species
    Mouse
  • Specificity
    Human and Bovine complex V beta subunit (ATPB).
  • Tested applications
    Suitable for: WB, IHC-Fr, ICC/IF, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Pig, Caenorhabditis elegans, Monkey
  • Immunogen

    Whole heart mitochondria (Human).

  • 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

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.

IHC-P Use at an assay dependent concentration.

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

  • 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 kDa



    Merged 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.

  • 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)
  • 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.
  • ab14730 (blue) at 1µg/ml staining ATPB in HL-60 cells and analyzed by Flow cytometry. Red histogram represents equal quantity of isotype control.
  • 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

  • ab14730 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.

    See Abreview

  • 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

References

This product has been referenced in:
See all 134 Publications for this product

Customer reviews and Q&As

1-6 of 6 Q&A

Answer

Thank you for contacting us.

The MSDS for these products have been added to each product datasheet. They should appear on the website within the next few hours (time for the cache to be regenerated).

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.



Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Answer

Thank you for your response and for your confirmation.

DISCOUNT CODE: 100%ABR-EXXXXX
Expiration date: 29/07/2012

I am very pleased to hear you would like to accept our offer and testab14730 in Pig WB. This code will give you 1 free PRIMARY ANTIBODY/PROTEIN before the expiration date. To redeem this offer, please submit an Abreview for in Pig WB and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you for your response.

Could you please confirm if you understand the Teams and Conditions? Your supervisor has called us previously for further explanation.

It would be much appreciated if you could also specify in what application you wish to test this antibody on pig samples?

I look forward to hearing from you soon.

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Answer

Thank you very much for your interest in ab14730 and for your recent phone call.

To our knowledge,ab14730 has not been tested in pig (Sus scrofa). Therefore, I can offer a discount off a future purchase if you buyab14730 now, test it inpig (Sus scrofa) and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free PRIMARY ANTIBODY.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and testab14730 in pig (Sus scrofa). I will then send a discount code. This code must be issued before purchasingab14730 so please wait for my reply before ordering.

2. Purchaseab14730 either by phone, fax, or online (www.abcam.com).

3. Test it in pig (Sus scrofa).

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any PRIMARY ANTIBODY ordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even ifab14730 turns out to be unsuitable for pig (Sus scrofa), you will still receive the discount on your next purchase after your Abreview has been submitted.

Could you please discuss this with your supervisor and let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Answer

Thank you for your enquiry and your interest in our products.

This antibody has been characterized for cross-reactivity with Mouse, Rat, Cow, Human, Caenorhabditis elegans, Monkey. Up till now, we have no data regarding pig (Sus scrofa).

Due to sequence homologyab14730 might detect porcine ATPB but we have no supporting evidence yet. You may wish to visit UniProt website for further information on sequence homology between different species:

http://www.uniprot.org/blast/uniprot/2012032260GH5D6JUV

If you wish to test this product in pig samples, I could offer you a testing discount. The Terms and Conditions of this offer can be found at: https://www.abcam.com/collaborationdiscount.

If you need any further assistance in the future, please do not hesitate to contact me.

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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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