RecombinantRabMAb

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

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Flow Cytometry (Intracellular) - Anti-TIM44 antibody [EPR16821-26] (ab201453)
  • Western blot - Anti-TIM44 antibody [EPR16821-26] (ab201453)
  • Western blot - Anti-TIM44 antibody [EPR16821-26] (ab201453)
  • Western blot - Anti-TIM44 antibody [EPR16821-26] (ab201453)
  • Immunocytochemistry/ Immunofluorescence - Anti-TIM44 antibody [EPR16821-26] (ab201453)
  • Immunocytochemistry/ Immunofluorescence - Anti-TIM44 antibody [EPR16821-26] (ab201453)
  • Immunoprecipitation - Anti-TIM44 antibody [EPR16821-26] (ab201453)
  • Anti-TIM44 antibody [EPR16821-26] (ab201453)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR16821-26] to TIM44
  • Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP
  • Reacts with: Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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 (Intra), WB, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Rat, Human

  • Immunogen

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

  • 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

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR16821-26

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

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 (Intra)
1/500.
WB (1)
1/2000. Detects a band of approximately 45 kDa (predicted molecular weight: 51 kDa).
ICC/IF
1/500.
IP
1/120.
Notes
Flow Cyt (Intra)
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 (Intracellular) - Anti-TIM44 antibody [EPR16821-26] (ab201453)
      Flow Cytometry (Intracellular) - 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.

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

    References (0)

    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.

    Customer reviews and Q&As

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    Western blot abreview for Anti-TIM44 antibody [EPR16821-26]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Cortical Neurons)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    25 µg
    Specification
    Cortical Neurons
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Read More

    Eyal Paz

    Verified customer

    Submitted Apr 06 2021

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