Recombinant
RabMAb

Anti-HCE antibody [EPR19384] (ab201046)

Overview

  • Product name
    Anti-HCE antibody [EPR19384]
    See all HCE primary antibodies
  • Description
    Rabbit monoclonal [EPR19384] to HCE
  • Tested applications
    Suitable for: IHC-P, WB, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human HCE aa 1-250. The exact sequence is proprietary.
    Database link: O60942

  • Positive control
    • WB: Human fetal liver and fetal kidney lysates; Ramos, HepG2, SH-SY5Y, HeLa, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain, kidney and spleen lysates; Rat brain and kidney lysates. IHC-P: Mouse cerebral cortex and rat spleen tissues. ICC/IF: HeLa and NIH/3T3 cells. IP: SH-SY5Y whole cell lysate.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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

Applications

Our Abpromise guarantee covers the use of ab201046 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
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

The IHC application is recommended for mouse and rat only.

WB 1/2000. Detects a band of approximately 69 kDa (predicted molecular weight: 69 kDa).
IP 1/50.
ICC/IF 1/500.

Target

  • Function
    Bifunctional mRNA-capping enzyme exhibiting RNA 5'-triphosphatase activity in the N-terminal part and mRNA guanylyltransferase activity in the C-terminal part. Catalyzes the first two steps of cap formation: by removing the gamma-phosphate from the 5'-triphosphate end of nascent mRNA to yield a diphosphate end, and by transferring the gmp moiety of GTP to the 5'-diphosphate terminus.
  • Tissue specificity
    Isoform 1 and isoform 4 (at a lesser extent) are expressed in cerebrum, cerebellum, thyroid, lung, heart, liver, kidney, spleen, large intestine, testis, skin and muscle.
  • Sequence similarities
    In the N-terminal section; belongs to the non-receptor class of the protein-tyrosine phosphatase family.
    In the C-terminal section; belongs to the eukaryotic GTase family.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • CAP1A antibody
    • GTase antibody
    • GTP--RNA guanylyltransferase antibody
    • hCAP antibody
    • HCAP1 antibody
    • HcE antibody
    • HCE1 antibody
    • MCE1 antibody
    • MCE1_HUMAN antibody
    • mRNA 5''-triphosphatase antibody
    • mRNA capping enzyme antibody
    • mRNA guanyltransferase antibody
    • mRNA guanylyltransferase antibody
    • Polynucleotide 5' trisphosphatase antibody
    • RNA guanylyltransferase and 5' phosphatase antibody
    • RNGTT antibody
    • TPase antibody
    see all

Images

  • All lanes : Anti-HCE antibody [EPR19384] (ab201046) at 1/2000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size : 69 kDa
    Observed band size : 69 kDa


    Exposure time : 3 minutes

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-HCE antibody [EPR19384] (ab201046) at 1/2000 dilution

    Lane 1 : Ramos (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 3 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
    Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size : 69 kDa
    Observed band size : 69 kDa


    Exposure time : 1 minute

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-HCE antibody [EPR19384] (ab201046) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat kidney lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size : 69 kDa
    Observed band size : 69 kDa


    Exposure time : 3 minutes

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-HCE antibody [EPR19384] (ab201046) at 1/2000 dilution

    Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size : 69 kDa
    Observed band size : 69 kDa


    Exposure time : 30 seconds

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling HCE with ab201046 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on neurons of mouse cerebral cortex is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling HCE with ab201046 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on lymphocytes of rat spleen is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling HCE with ab201046 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [EPR19384] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab201046 at 1/500 dilution followed by ab150120 at 1/1000 dilution.
    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling HCE with ab201046 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [EPR19384] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab201046 at 1/500 dilution followed by ab150120 at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • HCE was immunoprecipitated from 1mg of SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate with ab201046 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab201046 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.
    Lane 1: SH-SY5Y whole cell lysate 10µg (Input).
    Lane 2: ab201046 IP in SH-SY5Y whole cell lysate.
    Lane 3: Rabbit IgG,monoclonal [EPR19384] -Isotype Control (ab172730) instead of ab201046 in SH-SY5Y whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 30 seconds.

     

References

ab201046 has not yet been referenced specifically in any publications.

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