Overview

  • Product name

    Anti-LMO4 antibody [EPR6731(2)]
    See all LMO4 primary antibodies
  • Description

    Rabbit monoclonal [EPR6731(2)] to LMO4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: ICC or IHC-P
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human LMO4 (C terminal). The exact sequence is proprietary.

  • Positive control

    • 293T, Jurkat, SH-SY5Y, and NIH3T3 cell lysates, 293T cells
  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

     

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab131030 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. Predicted molecular weight: 17 kDa.
IP 1/20 - 1/100.
  • Application notes
    Is unsuitable for ICC or IHC-P.
  • Target

    Images

    • All lanes : Anti-LMO4 antibody [EPR6731(2)] (ab131030) at 1/1000 dilution

      Lane 1 : 293T cell lysate
      Lane 2 : Jurkat cell lysate
      Lane 3 : SH-SY5Y cell lysate
      Lane 4 : NIH3T3 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled Goat anti-Rabbit at 1/2000 dilution

      Predicted band size: 17 kDa



      Actual band size : 17 kDa
    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • All lanes : Anti-LMO4 antibody [EPR6731(2)] (ab131030) at 1/1000 dilution

      Lane 1 : 293T (human embryonic kidney) whole cell lysate
      Lane 2 : Jurkat (human acute T cell leukemia) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

      Predicted band size: 17 kDa
      Observed band size: 17 kDa



      Blocking/Diluting buffer 5% NFDM/TBST
    • All lanes : Anti-LMO4 antibody [EPR6731(2)] (ab131030) at 1/1000 dilution

      Lane 1 : NIH/3T3 (mouse embryo) whole cell lysate
      Lane 2 : Mouse cerebral cortex tissue lysate

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 17 kDa
      Observed band size: 17 kDa



      Blocking/Diluting buffer 5% NFDM/TBST
    • Purified ab131030 at 1/20 immunoprecipitating LMO4 in Jurkat whole cell lysate observed at 18 KDa (lanes 1 and 2).

      Lane 1 (input): Jurkat whole cell lysate 10ug

      Lane 2 (+): ab131030 + Jurkat whole cell lysate.

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab131030 in Jurkat whole cell lysate

      VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

      Blocking/Diluting buffer 5% NFDM/TBST.

    References

    ab131030 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Answer



    Protocol:


    Jurkat cell was used for the IP test. The concentration of the jurkat lysate was 3.5mg/ml. For the IP test, 6ul of the capture antibody was added to 300ul of the jurkat lysate (conduct on the ice).



    Protocol 1



    1. Thaw the IP lysates on the ice. Then centrifugate at 10,000 rpm for 2 min. to remove the cell fragments.

    * The protein concentration in the lysates is about 3.5mg/ml

    2. Add 6 ul of the capture antibody to 300 ul of the lysate (conduct on the ice). Swing slightly overnight at 4℃.

    3. Add 25 ul of the precooled beads. Swing slightly at 4 ℃ for 2 hours.

    4. Centrifuge at 10,000 rpm for 2 min.. Then wash the conjugate of beads-antibody-antigen with 1 ml of precooled TBS for 3 times.

    5. Re-suspend the conjugate of beads-antibody-antigen in 20 ul of 2×SDS loading buffer containing 5% beta-mercaptoethanol. Boil for 5min., and centrifuge at 10,000 rpm for 2 min..

    6. Load the supernatant on one lane, and apply on electrophoresis and electro-transformation.

    7. Treat the strip of membrane with 99.5% methanol for 15 sec.. Then rinse it on swing bed with deionized water for 5 min..

    8. Block the strip with 5% NFDM/TBST at RT for 1 hour on swing bed. Then wash it on swing bed with 1×TBST for 10 min..

    9. Incubate the strip with the blot antibody of 1:1000 dilution in 5% NFDM/TBST, at RT for 1 hour on swing bed.

    10. Wash the strip 3 times with 1×TBST on swing bed, 10 min. per time.

    11.Incubate with 2nd secondary antibody of 1:1000 dilution in 5% NFDM/TBST for 1 hour at RT on swing bed.

    12. Wash the strip 3 times with 1×TBST on swing bed, 10 min. per time.

    13. Add ECL substrate on the strip for 5 min.at RT. Then exposure for 10 sec. on X-ray film. (In order to get good image, the exposure time could be adjusted to 1 sec. to 30 sec.)

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