Recombinant
RabMAb

Recombinant Anti-TIM44 antibody [EPR16821-26] (ab201453)

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Overview

  • Product name

    Anti-TIM44 antibody [EPR16821-26]
    See all TIM44 primary antibodies

  • Description

    Rabbit monoclonal [EPR16821-26] to TIM44

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, WB, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Rat, Human

  • Immunogen

    Recombinant fragment within Human TIM44 aa 1-250. The exact sequence is proprietary.
    Database link: O43615

  • Positive control

    • WB: K562, Daudi, HepG2, PC-12 and C6 cell lysates; Human fetal liver lysate. ICC/IF: K562 and HepG2 cells. IP: K562 whole cell lysate.

  • General notes

     

     

    Previously labelled as TIMM44. 

    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

  • Form

    Liquid

  • Storage instructions

    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.

  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR16821-26

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab201453 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
Flow Cyt 1/500.
WB 1/2000. Detects a band of approximately 45 kDa (predicted molecular weight: 51 kDa).
ICC/IF 1/500.
IP 1/120.

Target

  • Relevance

    TIM44 is the essential component of the PAM complex, a complex required for the translocation of transit peptide-containing proteins from the inner membrane into the mitochondrial matrix in an ATP-dependent manner. It recruits mitochondrial HSP70 to drive protein translocation into the matrix using ATP as an energy source.

  • Cellular localization

    Mitochondrion inner membrane.

  • Database links

    • Alternative names

      • DKFZp686H05241 antibody
      • MIMT44 antibody
      • Mitochondrial inner membrane translocase antibody
      • TIM44 antibody
      • Translocase of inner mitochondrial membrane 44 antibody
      • Translocase of inner mitochondrial membrane 44 homolog (yeast) antibody
      see all

    Images

    • Flow Cytometry - Anti-TIM44 antibody [EPR16821-26] (ab201453)
      Flow Cytometry - Anti-TIM44 antibody [EPR16821-26] (ab201453)

      Flow Cytometry analysis of HepG2(human hepatocellular carcinoma) labelling TIM44 with purified ab201453 at 1/500 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Alexa Fluor® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

       

    • Western blot - Anti-TIM44 antibody [EPR16821-26] (ab201453)
      Western blot - Anti-TIM44 antibody [EPR16821-26] (ab201453)
      All lanes : Anti-TIM44 antibody [EPR16821-26] (ab201453) at 1/2000 dilution

      Lane 1 : K562 (Human chronic myelogenous leukemia cells from bone marrow) cell lysate
      Lane 2 : HepG2 (Human liver hepatocellular carcinoma) cell lysate
      Lane 3 : Daudi (Human Burkitt's lymphoma cell line) cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size: 51 kDa
      Observed band size: 45 kDa
      why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-TIM44 antibody [EPR16821-26] (ab201453)
      Western blot - Anti-TIM44 antibody [EPR16821-26] (ab201453)
      Anti-TIM44 antibody [EPR16821-26] (ab201453) at 1/10000 dilution + Human fetal liver lysate at 10 µg

      Secondary
      Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size: 51 kDa
      Observed band size: 45 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-TIM44 antibody [EPR16821-26] (ab201453)
      Western blot - Anti-TIM44 antibody [EPR16821-26] (ab201453)
      All lanes : Anti-TIM44 antibody [EPR16821-26] (ab201453) at 1/2000 dilution

      Lane 1 : C6 (Rat glial tumor cells) cell lysate
      Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size: 51 kDa
      Observed band size: 45 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence - Anti-TIM44 antibody [EPR16821-26] (ab201453)
      Immunocytochemistry/ Immunofluorescence - Anti-TIM44 antibody [EPR16821-26] (ab201453)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling TIM44 with ab201453 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasm staining on HepG2 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

      The negative controls are as follows:
      -ve control 1: ab201453 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
      -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

    • Immunocytochemistry/ Immunofluorescence - Anti-TIM44 antibody [EPR16821-26] (ab201453)
      Immunocytochemistry/ Immunofluorescence - Anti-TIM44 antibody [EPR16821-26] (ab201453)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling TIM44 with ab201453 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasm staining on K562 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

      The negative controls are as follows:
      -ve control 1: ab201453 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
      -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

    • Immunoprecipitation - Anti-TIM44 antibody [EPR16821-26] (ab201453)
      Immunoprecipitation - Anti-TIM44 antibody [EPR16821-26] (ab201453)

      TIM44 was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate with ab201453 at 1/120 dilution. Western blot was performed from the immunoprecipitate using ab201453 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution.

      Lane 1: K562 whole cell lysate 10 µg (Input). Lane 2: ab201453 IP in K562 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab201453 in K562 whole cell lysate.
      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 1 second.

    References

    Publishing research using ab201453? Please let us know so that we can cite the reference in this datasheet.

    ab201453 has not yet been referenced specifically in any publications.

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