Overview

  • Product name

    Anti-HTF9C/TRMT2A antibody [EPR16485-54]
    See all HTF9C/TRMT2A primary antibodies
  • Description

    Rabbit monoclonal [EPR16485-54] to HTF9C/TRMT2A
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human HTF9C/TRMT2A aa 100-400. The exact sequence is proprietary.
    Database link: Q8IZ69

  • Positive control

    • WB: Human fetal kidney lysate; MOLT-4, HEK293, Jurkat, HepG2, C6, PC-12, NIH/3T3 whole cell lysates. IHC-P: Human breast carcinoma tissue.
  • General notes

     

     

     This product was previously labelled as HTF9C

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab205616 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. Detects a band of approximately 75 kDa (predicted molecular weight: 69 kDa).
IP 1/100.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

Images

  • Anti-HTF9C/TRMT2A antibody [EPR16485-54] (ab205616) at 1/1000 dilution + Human fetal kidney at 20 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Developed using the ECL technique.

    Predicted band size: 69 kDa
    Observed band size: 75 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking and diluting buffer was 5% NFDM /TBST

  • All lanes : Anti-HTF9C/TRMT2A antibody [EPR16485-54] (ab205616) at 1/5000 dilution

    Lane 1 : MOLT-4 (Human lymphoblastic leukemia cell line) whole cell lysate
    Lane 2 : HEK293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 69 kDa
    Observed band size: 75 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking and diluting buffer was 5% NFDM /TBST

    Exposure time Lane 1 and 2: 10 seconds
    Lane 3: 1 minute

  • Anti-HTF9C/TRMT2A antibody [EPR16485-54] (ab205616) at 1/1000 dilution + HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 69 kDa
    Observed band size: 75 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking and diluting buffer was 5% NFDM /TBST

  • All lanes : Anti-HTF9C/TRMT2A antibody [EPR16485-54] (ab205616) at 1/1000 dilution

    Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 69 kDa
    Observed band size: 75 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking and diluting buffer was 5% NFDM /TBST

  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling HTF9C/TRMT2A with ab205616 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.

    Cytoplasmic staining on human breast carcinoma tissue is observed.

    Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody with secondary antibody, ab97051 at 1/500.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • HTF9C/TRMT2A was immunoprecipitated from 1mg of HEK293 (Human epithelial cell line from embryonic kidney) whole cell extract with ab205616 at 1/100.

    Western blot was performed from the immunoprecipitate using ab205616 at 1/5000 followed by VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/10000.

    Lane 1: HEK293 whole cell extract at 10ug

    Lane 2: HEK293 whole cell extract immunoprecipitated using ab205616

    Lane 3: HEK293 whole cell extract immunoprecipitated using ab172730 (Rabbit monoclonal IgG)

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

References

ab205616 has not yet been referenced specifically in any publications.

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