Recombinant
RabMAb

Recombinant Anti-Aldolase + Aldolase C antibody [EPR19355] (ab200049)

Overview

  • Product name

    Anti-Aldolase + Aldolase C antibody [EPR19355]
    See all Aldolase+Aldolase C primary antibodies
  • Description

    Rabbit monoclonal [EPR19355] to Aldolase + Aldolase C
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant full length protein within Human Aldolase aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P09972

  • Positive control

    • WB: Human Aldolase C full length recombinant protein; human Aldolase full length recombinant protein; HeLa, U-87 MG, C6, RAW 264.7 and NIH/3T3 whole cell lysates; human cerebellum, fetal liver, fetal heart, fetal kidney and fetal spleen lysates; rat brain and kidney lysates; mouse brain and heart lysates. ICC/IF: U-87 MG and NIH/3T3 cells. Flow Cyt: U-87 MG cells.
  • General notes

     

     

    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 ab200049 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 1/2000. Detects a band of approximately 39 kDa (predicted molecular weight: 39 kDa).
ICC/IF 1/500.
Flow Cyt 1/600.

Images

  • All lanes : Anti-Aldolase + Aldolase C antibody [EPR19355] (ab200049) at 1/1000 dilution

    Lane 1 : Human Aldolase C full length recombinant protein
    Lane 2 : Human Aldolase full length recombinant protein
    Lane 3 : Human Aldolase B full length recombinant protein

    Lysates/proteins at 0.01 µg per lane.

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

    Predicted band size: 39 kDa
    Observed band size: 39 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Human ALDOC full length recombinant protein contains aa2-364 with a His-Tag®. Human ALDOA and ALDOB full length recombinant protein contains aa1-364 with a His-Tag®. These three recombinant proteins were made in-house.

    This product showed very weak reactivity to Aldolase B, relative to Aldolase and Aldolase C, as demonstrated by WB of recombinant proteins.

  • Lanes 1-3 : Anti-Aldolase + Aldolase C antibody [EPR19355] (ab200049) at 1/5000 dilution
    Lanes 4-7 : Anti-Aldolase + Aldolase C antibody [EPR19355] (ab200049) at 1/2000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : U 87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate
    Lane 3 : Human cerebellum lysate
    Lane 4 : Human fetal liver lysate
    Lane 5 : Human fetal heart lysate
    Lane 6 : Human fetal kidney lysate
    Lane 7 : human fetal spleenlysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lanes 4-7 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 39 kDa
    Additional bands at: 39 kDa (possible non-specific secondary antibody binding)



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-3: 8 seconds; Lane 4-7: 10 seconds.

    The expression profile observed is consistent with what has been described in the literature (PMID:  24475166 and 17659271).

  • Lanes 1-6 : Anti-Aldolase + Aldolase C antibody [EPR19355] (ab200049) at 1/5000 dilution
    Lane 7 : Anti-Aldolase + Aldolase C antibody [EPR19355] (ab200049) at 1/2000 dilution

    Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 2 : Rat brain lysate
    Lane 3 : Rat kidney lysate
    Lane 4 : Mouse heart lysate
    Lane 5 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 6 : Mouse brain lysate
    Lane 7 : 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

    Predicted band size: 39 kDa
    Observed band size: 39 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-5: 8 seconds; Lane 6/7: 3 seconds.

     

  • Immunofluorescent analysis of 100% methanol-fixed U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling Aldolase C + Aldolase with ab200049 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on U-87 MG cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab200049 at 1/500 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunofluorescent analysis of 100% methanol-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Aldolase C + Aldolase with ab200049 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab200049 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling Aldolase + Aldolase C with ab200049 at 1/600 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.

References

ab200049 has not yet been referenced specifically in any publications.

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