Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ALK antibody [EPR24029-128] (ab275964)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR24029-128] to ALK
  • Suitable for: WB, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ALK antibody [EPR24029-128]
    See all ALK primary antibodies

  • Description

    Rabbit monoclonal [EPR24029-128] to ALK

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IP
    Mouse
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Mouse P5 brain tissue lysate; Rat P0 brain tissue lysate; Neuro-2a,SK-N-BE(2), IMR032 and SH-SY5Y whole cell lysates. IP: Neuro-2a and IMR-32 whole cell lysates.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

Images

  • Western blot - Anti-ALK antibody [EPR24029-128] (ab275964)
    Western blot - Anti-ALK antibody [EPR24029-128] (ab275964)
    All lanes : Anti-ALK antibody [EPR24029-128] (ab275964) at 1/1000 dilution

    Lane 1 : Mouse P5 brain tissue lysate
    Lane 2 : Mouse kidney tissue lysate
    Lane 3 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate
    Lane 4 : Rat P0 brain tissue lysate
    Lane 5 : Rat kidney tissue lysate

    Lysates/proteins at 40 µg per lane.

    Secondary
    All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

    Predicted band size: 176 kDa
    Observed band size: 140-250 kDa
    why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    ALK undergoes glycosylation and phosphorylation. ALK has both extracellular and intracellular cleavage sites. The molecular weight observed are consistent with what have been described in literatures (PMID: 30541790, 32344689).

    Negative control: kidney (PMID: 9053841).

    This blot was developed using a higher sensitivity ECL substrate.

    Exposure time: 3 minutes.

     

  • Immunoprecipitation - Anti-ALK antibody [EPR24029-128] (ab275964)
    Immunoprecipitation - Anti-ALK antibody [EPR24029-128] (ab275964)

    ALK was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate with ab275964 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275964 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/1000 dilution.

    Lane 1: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate 10 ug

    Lane 2: ab275964 IP in Neuro-2a whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275964 in Neuro-2a whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

     

  • Western blot - Anti-ALK antibody [EPR24029-128] (ab275964)
    Western blot - Anti-ALK antibody [EPR24029-128] (ab275964)
    All lanes : Anti-ALK antibody [EPR24029-128] (ab275964) at 1/1000 dilution

    Lane 1 : SK-N-BE(2) (human neuroblastoma neuroblast) whole cell lysate
    Lane 2 : IMR-32 (human neuroblastoma neuroblast) whole cell lysate
    Lane 3 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate
    Lane 4 : Raji (human burkitts lymphoma b lymphocyte) whole cell lysate
    Lane 5 : Ramos (human burkitts lymphoma b lymphocyte) whole cell lysate

    Lysates/proteins at 40 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

    Predicted band size: 176 kDa
    Observed band size: 140-250,70 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    ALK undergoes glycosylation and phosphorylation. ALK has both extracellular and intracellular cleavage sites. The 140-250 kDa bands observed are consistent with what have been described in literatures (PMID: 30541790, 32344689).

    A 70-kDa band is observed, it may correspond to N terminal truncated ALK (PMID: 30400214).

    Negative control: Raji and Ramos (PMID: 30541790).

    Exposure time: 26 seconds.

     

  • Immunoprecipitation - Anti-ALK antibody [EPR24029-128] (ab275964)
    Immunoprecipitation - Anti-ALK antibody [EPR24029-128] (ab275964)

    ALK was immunoprecipitated from 0.35 mg IMR-32 (human neuroblastoma neuroblast) whole cell lysate with ab275964 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275964 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: IMR-32 (human neuroblastoma neuroblast) whole cell lysate 10 ug

    Lane 2: ab275964 IP in IMR-32 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab275964 in IMR-32 whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 119 seconds.

     

  • Anti-ALK antibody [EPR24029-128] (ab275964)
    Anti-ALK antibody [EPR24029-128] (ab275964)

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