Anti-ATPB抗体[4.3E8.D10] - Mitochondrial Marker (ab5432)

Reviews (1)Q&A (7)References (13)

Key features and details

  • Mouse monoclonal [4.3E8.D10] to ATPB - Mitochondrial Marker
  • Suitable for: WB, ICC/IF, IP
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG1

概述

  • 产品名称

    Anti-ATPB抗体[4.3E8.D10] - Mitochondrial Marker
    参阅全部 ATPB 一抗

  • 描述

    小鼠单克隆抗体[4.3E8.D10] to ATPB - Mitochondrial Marker

  • 宿主

    Mouse

  • 特异性

    Detects the beta subunit of ATP synthase (ATPB) from mouse rat and human samples. This antibody is useful as a mitochondrial marker.

  • 经测试应用

    适用于: WB, ICC/IF, IPmore details

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Other Immunogen Type corresponding to ATPB. Intact rat mitochondria.

性能

应用

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.

应用 Ab评论 说明
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 57 kDa (predicted molecular weight: 57 kDa).
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.

靶标

  • 功能

    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.

  • 序列相似性

    Belongs to the ATPase alpha/beta chains family.

  • 细胞定位

    Mitochondrion. Mitochondrion inner membrane. Peripheral membrane protein.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • 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

图片

  • Western blot - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (ab5432)
    Western blot - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (ab5432)
    Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (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
  • Immunocytochemistry/ Immunofluorescence - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (ab5432)
    Immunocytochemistry/ Immunofluorescence - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (ab5432)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (ab5432)
    Immunocytochemistry/ Immunofluorescence - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (ab5432)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (ab5432)
    Immunocytochemistry/ Immunofluorescence - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (ab5432)

    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.

  • Immunoprecipitation - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (ab5432)
    Immunoprecipitation - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (ab5432)
    Immunoprecipatation of rat neuronal/glial cell extract using ab5432.
  • Immunocytochemistry/ Immunofluorescence - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (ab5432)
    Immunocytochemistry/ Immunofluorescence - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (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 - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (ab5432)
    Immunocytochemistry/ Immunofluorescence - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (ab5432)

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

文献 (13)

发表研究结果有使用 ab5432？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab5432 被引用在 13 文献中.

  • El-Khoury R  et al. Divergent Features of Mitochondrial Deficiencies in LGMD2A Associated With Novel Calpain-3 Mutations. J Neuropathol Exp Neurol 78:88-98 (2019). PubMed: 30500922
  • Tran L  et al. Mitochondrial ATP synthase ß-subunit production rate and ATP synthase specific activity are reduced in skeletal muscle of humans with obesity. Exp Physiol 104:126-135 (2019). PubMed: 30362197
  • 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). 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 . PubMed: 28005946
  • Jans DC  et al. STED super-resolution microscopy reveals an array of MINOS clusters along human mitochondria. Proc Natl Acad Sci U S A 110:8936-41 (2013). PubMed: 23676277
View all Publications for this product

客户评价及客户问答

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1-8 of 8 Abreviews or Q&A

Question

Thank you very much for the prompt response. I wonder of you also have human mitochondria similar to the bovine preparation? Also, please let me know if the anti ATP synthase that you sell (particularly the mouse monoclonal ab5432 and the monoclonal antobody clone 7E3F2 - AB110280) interact with the bovine mitochondria - AB110338. I thank you in advance for your answer as soon as you can.

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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

check whether there is any information on the immunogen of ab5432 or ab14730

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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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Western blot abreview for Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker

 Poor
Abreviews
Application
Western blot
Sample
Drosophila melanogaster Tissue lysate - whole (head)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
head
Blocking step
casein as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: 4°C
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Ms. Zhiyin Xun

Verified customer

提交于 Aug 14 2007

Question

I have a customer that is looking for an antibody that reacts to mitochondrial ATP synthase F1 complex, beta polypeptide (SwissProt P10719) for Western blotting. They would therefore like to know which domain was used as immunogen when this antibody was produced.

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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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Question

Q.1 Do you have any information about the epitope of these two antibodies? Mitochondrial ATP Synthase Subunit Beta antibody [3D5] (ab14730) ATP Synthase beta antibody [4.3E8.D10] (ab5432) I'd like to know which domain was used as immunogen when the antibody was produced. Q.2 What's the difference between these two antibodies? It seems that they have different application purposes.

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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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Question

I have searched the web of Abcam for antibodies detecting mitochondrial gene products. there are 30 antibodies for mitochondrial proteins, but they are not specifically marked as mitochondrial origin or nuclear origin. Could you help me to define which antibodies I can use for detecting the proteins from mitochondrial DNA? (the 13 proteins coded by mtDNA in mammals). Thank you very much for your help.

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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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Question

ANTIBODY CODE ab5432 DESCRIPTION OF THE PROBLEM No signal SAMPLE S.cereviciae/crude mitochondrial fraction PRIMARY ANTIBODY Ab cam/1:1000 dilution in blocking buffer, overnight incubation at 4C, washed 3 times for 10 min at RT. SECONDARY ANTIBODY Amersham/1:10000 dilution in blocking buffer/ overnight incubation at 4C/washed 3 times for 10 min at RT. DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED got positive signal against 'jurkat cell lysate (Santa Cruz Biotech)' ANTIBODY STORAGE CONDITIONS -20C, Aliquots were made. SAMPLE PREPARATION photassium phosphate buffer including protease inhibitor cocktail solution (sigma). haeted just before loaded. AMOUNT OF PROTEIN LOADED 50ug ELECTROPHORESIS/GEL CONDITIONS 10% SDS-PAGE TRANSFER AND BLOCKING CONDITIONS Tris/Glycine/20% MeOH, trasnfered ovrnight at constant 0.2A, 4C., 3% nonfat milk (Bio-rad) in TBS (0.05% Tween 20)

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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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Question

Customer's question: Is antibody ab5432 suitable for western blotting? How did you manage to seperate ATP synthase from the IGg bands in image 1633 if the it is 56 kDa?

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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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