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Tags & Cell Markers Subcellular Markers Plasma Membrane

Anti-Pma1抗体[40B7] (ab4645)

  • Datasheet
Reviews (1)Q&A (8)References (26)

Product price, shipping and contact information

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Key features and details

  • Mouse monoclonal [40B7] to Pma1
  • Suitable for: ICC/IF, WB
  • Reacts with: Saccharomyces cerevisiae
  • Isotype: IgG

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概述

  • 产品名称

    Anti-Pma1抗体[40B7]
  • 描述

    小鼠单克隆抗体[40B7] to Pma1
  • 宿主

    Mouse
  • 特异性

    Reacts with Pma1p based on the size of the band detected on western blots and the immunofluorescence localization pattern.
  • 经测试应用

    适用于: ICC/IF, WBmore details
  • 种属反应性

    与反应: Saccharomyces cerevisiae
  • 免疫原

    Tissue, cells or virus corresponding to Pma1. Yeast nuclear prep

  • 常规说明

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • 存储溶液

    Preservative: 0.065% Sodium azide
    Constituent: Tissue culture supernatant
  • Concentration information loading...
  • 纯度

    Tissue culture supernatant
  • 纯化说明

    Sterile filtered.
  • 克隆

    单克隆
  • 克隆编号

    40B7
  • 同种型

    IgG
  • 研究领域

    • Tags & Cell Markers
    • Subcellular Markers
    • Plasma Membrane

相关产品

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG - Isotype Control (ab37355)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab4645于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ICC/IF
Use at an assay dependent concentration.
WB (1)
Use at an assay dependent concentration.

(cell lysates, ECL detection). For non-ECL western detection methods, 1/1000 - 1/5000. Also detects Pma1p-containing fusion proteins on western blots.

说明
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.

(cell lysates, ECL detection). For non-ECL western detection methods, 1/1000 - 1/5000. Also detects Pma1p-containing fusion proteins on western blots.

靶标

  • 相关性

    Pma1p is a plasma membrane ATPase.
  • 数据库链接

    • Entrez Gene: 852876 Saccharomyces cerevisiae
    • SwissProt: P05030 Saccharomyces cerevisiae
    • 别名

      • Plasma membrane ATPase 1 antibody
      • Plasma membrane H+ ATPase antibody
      • Pma 1 antibody
      • Pma1p antibody
      • Proton pump 1 antibody
      see all

    实验方案

    • Western blot protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    数据表及文件

    • Datasheet download

      Download

    文献 (26)

    发表研究结果有使用 ab4645?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab4645 被引用在 26 文献中.

    • Gersing SK  et al. Mapping the degradation pathway of a disease-linked aspartoacylase variant. PLoS Genet 17:e1009539 (2021). PubMed: 33914734
    • Nightingale DJH  et al. A Protocol to Map the Spatial Proteome Using HyperLOPIT in Saccharomyces cerevisiae. Bio Protoc 9:e3303 (2019). PubMed: 33654815
    • Tremblay-Laganière C  et al. Polychlorinated biphenyl 126 exposure in rats alters skeletal muscle mitochondrial function. Environ Sci Pollut Res Int 26:2375-2386 (2019). PubMed: 30467749
    • Locke MN & Thorner J Rab5 GTPases are required for optimal TORC2 function. J Cell Biol 218:961-976 (2019). PubMed: 30578283
    • Nightingale DJH  et al. Mapping the Saccharomyces cerevisiae Spatial Proteome with High Resolution Using hyperLOPIT. Methods Mol Biol 2049:165-190 (2019). PubMed: 31602611
    View all Publications for this product

    客户评价及客户问答

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    提交评价 提交问题

    1-9 of 9 Abreviews or Q&A

    Western blot abreview for Anti-Pma1 antibody [40B7] - Plasma Membrane Marker

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Saccharomyces cerevisiae Cell lysate - other (Yeast membrane preparation)
    Loading amount
    5 µg
    Specification
    Yeast membrane preparation
    Gel Running Conditions
    Reduced Denaturing (7.5%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    提交于 Aug 07 2009

    Question

    Thank you for the reply. Regarding your questions:

    1) Was the transfer to PVDF verified with a Ponceau stain or another method?

    A: I did not Ponceau stain, however the blot was cut in half and the bottom section (approximately 60kD to the bottom) was probed for Pgk1 as a loading control, which showed that there was abundant protein transferred.

    2) Were the secondary antibodies anti-mouse IgG or anti-mouse IgM?

    A: The secondary's used (Goat Anti-Mouse and Donkey Anti-Mouse) were both IgG's (Heavy and Light Chain). I saw that the Pma1 antibody is an IgM antibody, but my understanding is that it should still react with IgG (H+L) secondary ab's, as it would react with the approriate light chain.

    3) What kind of samples you using and what species are they from?

    A: The samples ran ranged from both vegetative and sporulation protein extracts (extracted using alkaline lysis followed by TCA precipitation) as well as crude extracts from subcellular fractionation (supernatant and pellets from 300g, 14,000g, and 100,000g). All samples were from S. cerevisiae.

    I will certainly try both recommendations you had--increasing the protein loaded as well as antibody concentration. How high would you recommend increasing the antibody? My primary concern is that under the conditions I have used I cannot even see a faint band whatsoever. Would you believe that the inability to detect Pma1 had anything to do with solublization?

    Thank you for Abcam's product guarantee regarding replacement, credit, or refund. I have been very satisfied with the other antibodies we purchased from Abcam, and will discuss these options with my PI.

    Thank you for your speedy response,

    Read More

    Abcam community

    Verified customer

    Asked on Oct 19 2012

    Answer

    Thank you very much for your reply.

    The anti-IgG (H&L) secondary will bind to the conserved light chains of the IgM primary. I suggest using the secondary at a higher concentration than you normally would with an IgG primary, but I'd expect this secondary to be fine. You can try increasing the primary to 1:500, and the secondary to 1:5000 to see if this helps.

    To confirm solubility of membrane proteins you could try probing for another membrane marker, if you have an antibody to a different target available. I'm not sure of a better way to confirm this, but if the proteins didn't fully extract then this could be the cause of no signal. We do have one other antibody for an S. cerevisiae membrane protein, ab110326-

    https://www.abcam.com/VDACPorin-antibody-16G9E6BC4-Mitochondrial-Loading-Control-ab110326.html

    Would this be helpful for you? I look forward to hearing from you after discussing with your PI. Please let me know if you have any questions or if there is anything else that we can do for you. Have a nice weekend!

    Read More

    Abcam Scientific Support

    回复于 Oct 19 2012

    Question


    Inquiry: We purchased the Pma1 antibody for use in subcellular fractionation but have been unable to get any positive signal when used with both protein extracts from the fractionation procedure, as well as from protein extracts from a standard TCA precipitation. Amount of protein loaded: Varied levels from low to high amounts (high equates to ˜2.5OD600 from a TCA prep) Blocking conditions: 60min at room temp with agitation in Odyssey Blocking Buffer Detection method: Odyssey IR Fluorescence Detection Electrophoresis and transfer conditions: Used 7%, 10%, and 12% gels. Wet transfer, 200mA for 90min onto Low-Fluorescence PVDF. Primary antibody conditions (diluent/dilution, incubation time/temperature): 1:1,000 in Odyssey Blocking Buffer. Have done incubations at both 1hr room temp with agitation, as well as 4C with agitation overnight. Secondary antibody conditions (diluent/dilution, incubation time/temperature): 1:10,000 of IR Dye Goat-Anti-Mouse in Odyssey Blocking Buffer (also tried 1:10,000 of IR Dye Donkey Anti-Mouse). Have tried 1hr, 2hrs, and 4hrs of secondary incubation at room temp with agitation. Wash steps: Varied from gentle washing (1xPBST 3x5min after primary and then again after secondary) to more stringent washing (1XPBST 4x5min subsequent to primary and secondary incubations, with the secondary incubation being followed by a final 1XPBS rinse)

    Read More

    Abcam community

    Verified customer

    Asked on Oct 18 2012

    Answer

    Thank you very much for contacting us and letting us know about the trouble with this antibody.

    Thank you also for sending so many details about your protocol. I do have a couple of questions, just so I can further understand the situation:

    1) Was the transfer to PVDF verified with a Ponceau stain or another method?
    2) Were the secondary antibodies anti-mouse IgG or anti-mouse IgM?
    3) What kind of samples you using and what species are they from?

    I would expect the antibody to work under the conditions you've described, but it may be helpful to load more protein and increase the concentration of antibody. I understand that you have already tried some troubleshooting, and this antibody is guaranteed to work in WB with S. cerevisiae samples, so if you would like a replacement, credit, or refund, I'll be happy to set that up for you.

    I look forward to hearing from you. If you have any further questions or need anything else, please let me know and I'll be happy to help.

    Read More

    Abcam Scientific Support

    回复于 Oct 18 2012

    Question

    We bought the anti-Pma1 antidody (ab4645)
    Purcchase order and we couldn't see any band at 100KDA could
    we have a replacement for an other antibody????
    You will find attached the blot and the conditions used are below.
    Sample A2780 Human ovarian cell line
    Loading amount: 20 µg
    Gel Running Conditions: 12%
    Blocking step: Milk as blocking agent for 1 hour - Concentration: 5
    % - Temperature: 25°C
    Dilution 1/7000
    Incubation time: O/N Temperature: 4°C Diluent: PBS+tween
    Secondary antibody
    Name: anti-mouse IgM
    Dilution: 1/3000
    Detection
    Detection method: West Femto
    Exposure: 2 minutes
    Bands: NO BAND AT 100 kDa!!!!! only Non-specific bands!!!!!!

    Read More

    Abcam community

    Verified customer

    Asked on Jul 20 2012

    Answer

    Thank you for contacting Abcam.

    Under our Abpromise, we guarantee this antibody to work in western on yeast samples. As we had not tested in oh human samples, then we could not guarantee that it would work on this species.

    I would be very happy to help you troubleshoot though and see if we can try and get the antibody working. A few protocol variations that may help include:

    1 - Block in BSA, as this would help to reduce background and allow a longer exposure to see the 100kDa bands, if the antibody detects human samples.

    2 - Increase the total amount of protein loaded to 40ug.

    3 - Increase the primary antibody concentration from 1/7000, to 1/1000.

    If the above protocol variations do not prove to be successful, then please let me know and we can try and come up with other solutions.

    Also for your future reference, if you would ever like to use on of our antibodies in an untested application or species, then you could take advantage of our testing discount program:

    https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount



    Please let me know if there is anything else I can help you with.

    Read More

    Abcam Scientific Support

    回复于 Jul 20 2012

    Question

    Hi, Could you recommend a good antibody that I could use as loading control for western blot on cell membrane fractions? Thanks

    Read More

    Abcam community

    Verified customer

    Asked on Mar 30 2012

    Answer

    Thank you for your enquiry.

    Below is a selection of antibodies that are suitable for detecting the plasma membrane and nuclear envelope. They all work in western blot and I would review the datasheets to confirm which species they are tested against.

    Plasma membrane
    ab4645 Anti-Pma1 antibody [40B7]
    www.abcam.com/4645

    ab7671 Anti-alpha 1 Sodium Potassium ATPase antibody [464.6]
    www.abcam.com/ab7671

    ab16505 Anti-pan Cadherin antibody
    www.abcam.com/ab16505

    ab22744 Anti-pan Cadherin antibody [mAbcam22744]
    www.abcam.com/ab22744

    Nuclear envelope
    ab8980 Anti-Lamin A antibody [133A2]
    www.abcam.com/ab8980

    ab8984 Anti-Lamin A + C antibody [131C3]
    www.abcam.com/ab8984

    ab16048 Anti-Lamin B1 antibody
    www.abcam.com/ab16048

    I hope this is useful. Just let me know if you have further questions.

    Read More

    Abcam Scientific Support

    回复于 Mar 30 2012

    Question

    Bonjour je cherche un anticorps dirigé contre la membrane du reticulum endoplasmique de sacchaormyces cerevisae (ou pichia pastoris) Avez vous ce produit?

    Read More

    Abcam community

    Verified customer

    Asked on Sep 15 2011

    Answer

    Merci beaucoup pour votre intérêt pour nos anticorps. Nous avons trois anticorps actuellement qui pourront correspondre a vos besoin. Veuillez vérifier leur fiche technique s'il vous plaît. Je n'ai pas trouvé un anticorps testé contre pichia pastoris par contre. Dans quels applications est-ce que vous avez besoin d'utiliser cet anticorps? Anti-PDI antibody [PDI-38H8] - ER Marker (ab30319) Click here (or use the following: https://www.abcam.com/P4HB-antibody-PDI-38H8-ab30319.html). ti-Ribophorin I antibody (ab52672) Click here (or use the following: https://www.abcam.com/Ribophorin-I-antibody-ab52672.html). Anti-KDEL antibody [10C3] (ab94746) Click here (or use the following: https://www.abcam.com/HUS1B-antibody-ab4746.html). J'espère que ces informations vous sont utiles et que un des anticorps ci dessus peux correspondre à vos besoins. N'hésitez pas à nous recontacter si vous avez des autres questions.

    Read More

    Abcam Scientific Support

    回复于 Sep 15 2011

    Question

    What is the isotype of this antibody?

    Read More

    Abcam community

    Verified customer

    Asked on Apr 10 2006

    Answer

    Thank you for your enquiry. The isotype of this antibody is IgG3 with kappa light chains. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    回复于 Apr 24 2006

    Question

    Does ab4645 interact with the Canida albican homolog of PMA1?

    Read More

    Abcam community

    Verified customer

    Asked on Aug 12 2004

    Answer

    Thank you for your enquiry. All the information we have on species cross reactivity is specified on the datasheet, these are updated as soon as any new information is brought to our attention. As far as we are aware, cross reactivity with Canida albican has not yet been tested for use with ab4645. Should you decide to go ahead and purchase this product, please let us know how you get on and in return we will forward a reward of your choice, typically an Amazon gift voucher. Please let me know if you require any further assistance.

    Read More

    Abcam Scientific Support

    回复于 Aug 12 2004

    Question

    Customer's question: Regarding ab4645, I would need to know the epitope to which this antibody has been produced.

    Read More

    Abcam community

    Verified customer

    Asked on May 03 2004

    Answer

    The Pma1p protein is very antigenic, and antibodies to it turn up frequently if yeast membrane preps are used as the immunogen. However, what the exact antigenic region is, is not currently known.

    Read More

    Abcam Scientific Support

    回复于 May 11 2004

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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