Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-PFKFB3 antibody [EPR12594] - BSA and Azide free (ab218121)

Overview

  • Product name

    Anti-PFKFB3 antibody [EPR12594] - BSA and Azide free
    See all PFKFB3 primary antibodies
  • Description

    Rabbit monoclonal [EPR12594] to PFKFB3 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, ICC/IF, WB, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment aa 300 to the C-terminus. The exact sequence is proprietary.
    Database link: Q16875
    (Peptide available as ab192017)

  • Positive control

    • Jurkat and HeLa whole cell lysate (ab150035); Human melanoma tissue; HeLa and A431 cells.
  • General notes

    Ab218121 is the carrier-free version of ab181861. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab218121 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 ab218121 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
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 58 kDa (predicted molecular weight: 60 kDa).
IHC-P Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

Target

  • Function

    Synthesis and degradation of fructose 2,6-bisphosphate.
  • Tissue specificity

    Ubiquitous.
  • Sequence similarities

    In the C-terminal section; belongs to the phosphoglycerate mutase family.
  • Post-translational
    modifications

    Phosphorylation by AMPK stimulates activity.
  • Information by UniProt
  • Database links

  • Alternative names

    • 6 phosphofructo 2 kinase/ fructose 2,6 bisphosphatase antibody
    • 6 phosphofructo 2 kinase/fructose 2,6 biphosphatase 3 antibody
    • 6-bisphosphatase antibody
    • 6-P2ase 3 antibody
    • 6-P2ASE brain/placenta-type isozyme antibody
    • 6PF 2 K/Fru 2,6 P2ASE brain/placenta type isozyme antibody
    • 6PF 2-K/Fru 2,6 P2ase 3 antibody
    • 6PF-2-K/Fru-2 antibody
    • F263_HUMAN antibody
    • fructose 6 phosphate,2 kinase/fructose 2, 6 bisphosphatase antibody
    • Fructose-2 antibody
    • Inducible 6 phosphofructo 2 kinase/fructose 2,6 bisphosphatase antibody
    • iPFK 2 antibody
    • iPFK-2 antibody
    • IPFK2 antibody
    • PFK/FBPase 3 antibody
    • PFK2 antibody
    • PFKFB3 antibody
    • Renal carcinoma antigen NY REN 56 antibody
    • Renal carcinoma antigen NY-REN-56 antibody
    • uPFK antibody
    see all

Images

  • This WB data was generated using the same anti-PFKFB3 antibody clone, EPR12594, in a different buffer formulation (cat# ab181861).

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: PFKFB3 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: Jurkat whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab181861 observed at 60 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab181861 was shown to specifically react with PFKFB3 in wild-type HAP1 cells as signal was lost in PFKFB3 knockout cells. Wild-type and PFKFB3 knockout samples were subjected to SDS-PAGE. Ab181861 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry analysis of A431(human epidermoid carcinoma) cells labeling PFKFB3 with purified ab181861 at 1/210 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181861).

  • Western blot analysis of PFKFB3 in HeLa cell lysate immunoprecipitated using ab181861 at 1/50 dilution (Lane 1). Lane 2: Negative control.

    Secondary antibody: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181861).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed A431 cells labeling PFKFB3 with ab181861 at 1/100 dilution, followed by Goat anti rabbit IgG (Alexa Fluor®555) at 1/200 dilution. Counter stained with Dapi (blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181861).

  • Immunofluorescent analysis of acetone-fixed HeLa cells labeling PFKFB3 with ab181861 at 1/100 dilution, followed by Goat anti rabbit IgG (Alexa Fluor®555) at 1/200 dilution. Counter stained with Dapi (blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181861).

  • This IHC data was generated using the same anti-PFKFB3 antibody clone, EPR12594, in a different buffer formulation (cat# ab181861).

    Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling PFKFB3 with ab181861 at 1/50 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

References

This product has been referenced in:

  • Zhang J  et al. Protein kinase D3 promotes gastric cancer development through p65/6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 activation of glycolysis. Exp Cell Res 380:188-197 (2019). Read more (PubMed: 31026442) »
  • Jiang H  et al. PFKFB3-Driven Macrophage Glycolytic Metabolism Is a Crucial Component of Innate Antiviral Defense. J Immunol 197:2880-90 (2016). Read more (PubMed: 27566823) »
See all 2 Publications for this product

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