Overview

  • Product name

    Anti-VPS34 antibody [EPR5301]
    See all VPS34 primary antibodies
  • Description

    Rabbit monoclonal [EPR5301] to VPS34
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human VPS34 aa 800-900. The exact sequence is proprietary.

  • Positive control

    • L6, C2C12, 293T, SHYSY-5Y cell lysates; Mouse brain tissue lysate; Human gastric adenocarcinoma tissue
  • General notes

     

     

     This product was previously labelled as PI 3 Kinase Class 3

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab124905 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 1/1000 - 1/10000. Detects a band of approximately 100 kDa (predicted molecular weight: 102 kDa).
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt,ICC or IP.
  • Target

    • Function

      Catalytic subunit of the PI3K complex that mediates formation of phosphatidylinositol 3-phosphate. Involved in the transport of lysosomal enzyme precursors to lysosomes. Required for the abcission step in cytokinesis.
    • Tissue specificity

      Ubiquitously expressed, with a highest expression in skeletal muscle.
    • Sequence similarities

      Belongs to the PI3/PI4-kinase family.
      Contains 1 PI3K/PI4K domain.
    • Cellular localization

      Midbody.
    • Information by UniProt
    • Database links

    • Alternative names

      • hVps34 antibody
      • MGC61518 antibody
      • Phosphatidylinositol 3 kinase catalytic subunit type 3 antibody
      • Phosphatidylinositol 3 kinase class 3 antibody
      • Phosphatidylinositol 3 kinase p100 subunit antibody
      • Phosphatidylinositol 3-kinase catalytic subunit type 3 antibody
      • Phosphatidylinositol 3-kinase p100 subunit antibody
      • Phosphoinositide 3 kinase class 3 antibody
      • Phosphoinositide-3-kinase class 3 antibody
      • PI3 kinase type 3 antibody
      • PI3-kinase type 3 antibody
      • PI3K type 3 antibody
      • Pik3c3 antibody
      • PK3C3_HUMAN antibody
      • PtdIns 3 kinase type 3 antibody
      • PtdIns-3-kinase type 3 antibody
      • Vps 34 antibody
      • Vps34 antibody
      see all

    Images

    • All lanes : Anti-VPS34 antibody [EPR5301] (ab124905) at 1/1000 dilution

      Lane 1 : L6 cell lysate
      Lane 2 : C2C12 cell lysate
      Lane 3 : 293T cell lysate
      Lane 4 : Mouse brain tissue lysate
      Lane 5 : SHSY-5Y cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 102 kDa
      Observed band size: 100 kDa
      why is the actual band size different from the predicted?

    • ab124905, at 1/50 dilution, staining PI 3 Kinase in paraffin-embedded Human gastric adenocarcinoma tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    This product has been referenced in:

    See all 8 Publications for this product

    Customer reviews and Q&As

    1-4 of 4 Abreviews or Q&A

    Answer

    Thank you for your reply. Kate is currently away from the office but I will try to help as best I can in this case.

    Unfortunately, we do not currently have any alternative lots of this antibody in stock, however your customer may want to consider ab73262, or I can provide an alternative vial of ab124905.

    I look forward to receiving your reply.

    Read More

    Answer

    Thank you for your message and for providing kindly this further information.

    I appreciate the time you have spent on these experiments and it is regrettable the results have not been successful. I would be pleased to arrange a refund or credit note in compensation.

    In addition to this, experiments can often require individual optimization, andI suggest it would be beneficial to consider trying RIPA lysis buffer and a higher concentration of antibody for future experiments. Optimization such as this can sometimes help to improve results.

    I look forward to hearing from you with details of how you would like to proceed in this case.

    Read More

    Question

    Product code: 124905
    Lot number: GR78448-2

    Inquiry: Antibody code: ab124905
    Batch number: GR78448-2
    Antibody storage conditions (temperature/reconstitution etc): -20oC
    Description of the problem (high background, wrong band size, more bands, no band etc.):
    No bands
    Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.):
    total cellular proteins purified from Emiliania huxleyi, proteins isolated from human fibroblasts cells that strongly express the protein were used as positive control (other tests on this sample showed that it indeed possesses the protein). No bands were detected in the positive control.
    Sample preparation (Buffer/Protease inhibitors/Heating sample etc.): Tris pH=8, EDTA, SDS, Protease inhibitor.
    Amount of protein loaded: 30ug
    Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.): Reducing 4-16% precast Criterion TGX gel (BioRad)
    Transfer and blocking conditions (Buffer/time period, Blocking agent etc.): As described by BioRad. Identical gels were used for WB with a different antibody (ab4753) that worked well
    Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step): ab124905 1:2000
    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step): Goat anti Rabbit HRP (Jackson 111-035-003)
    Detection method (ECL, ECLPlus etc.): ECL Plus Positive and negative controls used (please specify):
    Positive control – proteins isolated from human fibroblasts cells that strongly express the protein were used as positive control (other tests on this sample showed that it indeed possesses the protein).
    OPTIMIZATION ATTEMPTS (PROBLEM SOLVING)
    How many times have you tried the Western? 3
    Have you run a "No Primary" control? yes
    Do you obtain the same results every time? e.g. are the background bands always in the same place? yes
    What steps have you altered? Increasing concentration of primary antibody, Increasing incubation time with primary antibody, increasing protein concentration on the gel.

    Read More
    Answer

    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

    The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

    I would like to reassure you that ab124905 is tested and covered by our 6 month guarantee for use in WB and human samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

    Reviewing this case, I would like to offer some suggestions to help optimise the results. I would also appreciate if you can confirm some further details:

    1. I can suggest to try RIPA rather than Tris buffer to lyse the samples. This may provide a more suitable protein preparation.

    2. Try the antibody at a higher concentration to increase the signal, for example 1:1000 or even 1:500. Incubating overnight at4oC can often provide more specific and efficient staining.

    3. Has the transfer to the membrane and quality of the sample been assessed using a loading control?

    4. Is the current vial of secondary antibody working well with other primary antibodies?

    I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

    Read More

    Answer

    Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

    The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

    Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial. I have therefore agreed, an asked our Finance department to issue a credit note for you.




    If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

    Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

    The credit note ID is for your reference only and does not automatically guarantee the credit.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

    Read More

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