Overview

  • Product name

    Anti-DRAM antibody [M3-P4B4]
    See all DRAM primary antibodies
  • Description

    Mouse monoclonal [M3-P4B4] to DRAM
  • Host species

    Mouse
  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human DRAM aa 150-250.

  • Positive control

    • This antibody gave a positive signal in HeLa whole cell lysate.
  • 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 ab64739 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 10 µg/ml. Predicted molecular weight: 26 kDa.
  • Application notes
    Is unsuitable for ICC/IF.
  • Target

    • Function

      Lysosomal modulator of autophagy that plays a central role in p53/TP53-mediated apoptosis. Not involved in p73/TP73-mediated autophagy.
    • Sequence similarities

      Belongs to the DRAM/TMEM150 family.
    • Cellular localization

      Lysosome membrane.
    • Information by UniProt
    • Database links

    • Alternative names

      • Damage regulated autophagy modulator antibody
      • Damage-regulated autophagy modulator 1 antibody
      • Damage-regulated autophagy modulator antibody
      • DNA damage-regulated autophagy modulator protein 1 antibody
      • DRAM antibody
      • dram1 antibody
      • DRAM1_HUMAN antibody
      • FLJ11259 antibody
      see all

    Images

    • Anti-DRAM antibody [M3-P4B4] (ab64739) at 10 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 20 µg

      Secondary
      Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Performed under reducing conditions.

      Predicted band size: 26 kDa
      Observed band size: 26 kDa


      Exposure time: 20 minutes

    References

    ab64739 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    1-4 of 4 Abreviews or Q&A

    Answer

    Thank you very much for confirming the details.

    I have checked the history of this antibody and can confirm that we haven't received specificity complaints for this product so I am sure protocol troubleshooting will help in getting a clean western blot results. I have following suggestions;

    - - In our own lab we test both milk and BSA as blocking solution. 5% BSA, 2 hours incubation time is the most common method and gives excellent results
    - We heat the lysates for 10 minutes at 100C
    - No primary control; this helps checking the non specificity due to secondary antibody
    - The problem might be with the lysates preparation, I would recommend using NP-40 or nonidet-P40 buffer for cell lysis. The ingredients can be found by clicking the following link;

    https://www.abcam.com/index.html?pageconfig=resource&rid=11414

    - For best results we recommend customers adding protease inhibitors. If possible, please add these in lysis buffer.
    - Positive control; Hela cells is the best positive control to be used with this antibody

    I hope the antibody would work following these suggestions. If in case the antibody fails again please contact me again for further assistance.

    Read More

    Answer

    Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

    I have read the details you have kindly provided and would like to ask few questions in order to understand the problem better;

    - What is the purchase order number of the product and how it was stored?
    - What is the lysate type and species and how it was prepared?
    - Have you added protease inhibitors? Was the lysate ice cold?
    - The Santa Cruz antibody also shows a large array of bands and no bands with Abcam ab, might be indicating the lysate problem. Have you used similar lysate preparation procedure successfully before?
    - How long the lysates were heated in sample buffer?
    - Could you provide the species of secondary antibody?

    I am looking forward to hearing from you soon.

    Read More
    This product is known to not work in this application or species.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (glioma stem cells)
    Specification
    glioma stem cells
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - triton-x 0.1%
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

    Abcam user community

    Verified customer

    Submitted Apr 26 2012

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (HeLa Ms Stomach Ms Kidney Ms Liver)
    Loading amount
    20 µg
    Specification
    HeLa Ms Stomach Ms Kidney Ms Liver
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C

    Mr. Antibody Solutions

    Verified customer

    Submitted May 25 2010

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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