Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-PFKFB3 antibody [EPR12594] (ab181861)

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Overview

  • Product name

    Anti-PFKFB3 antibody [EPR12594]
    See all PFKFB3 primary antibodies

  • Description

    Rabbit monoclonal [EPR12594] to PFKFB3

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human PFKFB3 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

    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

Applications

Our Abpromise guarantee covers the use of ab181861 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 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 60 kDa).
IP 1/50.
ICC/IF 1/100.
IHC-P 1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.

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

    see all

  • 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

  • Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861)
    Western blot - Anti-PFKFB3 antibody [EPR12594] (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.

  • Immunocytochemistry/ Immunofluorescence - Anti-PFKFB3 antibody [EPR12594] (ab181861)
    Immunocytochemistry/ Immunofluorescence - Anti-PFKFB3 antibody [EPR12594] (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).

  • Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861)
    Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861)
    All lanes : Anti-PFKFB3 antibody [EPR12594] (ab181861) at 1/20000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

    Predicted band size: 60 kDa
    Observed band size: 58 kDa
    why is the actual band size different from the predicted?



    Blocking buffer:  5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PFKFB3 antibody [EPR12594] (ab181861)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PFKFB3 antibody [EPR12594] (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.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-PFKFB3 antibody [EPR12594] (ab181861)
    Flow Cytometry - Anti-PFKFB3 antibody [EPR12594] (ab181861)

    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.

  • Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (ab181861)
    Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (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.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-PFKFB3 antibody [EPR12594] (ab181861)
    Immunocytochemistry/ Immunofluorescence - Anti-PFKFB3 antibody [EPR12594] (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).

References

Publishing research using ab181861? Please let us know so that we can cite the reference in this datasheet.

ab181861 has been referenced in 9 publications.

  • Zhang X  et al. Circular RNA circNRIP1 acts as a microRNA-149-5p sponge to promote gastric cancer progression via the AKT1/mTOR pathway. Mol Cancer 18:20 (2019). PubMed: 30717751
  • Wu Y  et al. Effect of microRNA-26a on vascular endothelial cell injury caused by lower extremity ischemia-reperfusion injury through the AMPK pathway by targeting PFKFB3. J Cell Physiol 234:2916-2928 (2019). PubMed: 30132885
  • Montemurro C  et al. IAPP toxicity activates HIF1a/PFKFB3 signaling delaying ß-cell loss at the expense of ß-cell function. Nat Commun 10:2679 (2019). PubMed: 31213603
  • Zhang XC  et al. YY1/LncRNA GAS5 complex aggravates cerebral ischemia/reperfusion injury through enhancing neuronal glycolysis. Neuropharmacology 158:107682 (2019). PubMed: 31278927
  • Xu C  et al. Canagliflozin exerts anti-inflammatory effects by inhibiting intracellular glucose metabolism and promoting autophagy in immune cells. Biochem Pharmacol 152:45-59 (2018). WB ; Mouse . PubMed: 29551587
  • Yan S  et al. 6-Phosphofructo-2-kinase/fructose-2,6-bisphosphatase isoform 3 spatially mediates autophagy through the AMPK signaling pathway. Oncotarget 8:80909-80922 (2017). PubMed: 29113354
  • He X  et al. Endothelial specific SIRT3 deletion impairs glycolysis and angiogenesis and causes diastolic dysfunction. J Mol Cell Cardiol 112:104-113 (2017). PubMed: 28935506
  • Jiang H  et al. PFKFB3-Driven Macrophage Glycolytic Metabolism Is a Crucial Component of Innate Antiviral Defense. J Immunol 197:2880-90 (2016). PubMed: 27566823
  • Lu Q  et al. Akt inhibition attenuates rasfonin-induced autophagy and apoptosis through the glycolytic pathway in renal cancer cells. Cell Death Dis 6:e2005 (2015). Human . PubMed: 26633711

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Western blot abreview for Anti-PFKFB3 antibody [EPR12594]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Murine breast cancer cells)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
5 µg
Specification
Murine breast cancer cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C
Read More

Alyssa La Belle

Verified customer

Submitted May 21 2019

Western blot abreview for Anti-PFKFB3 antibody [EPR12594]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Cow Cell lysate - whole cell (VVEC)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
20 µg
Treatment
100 uM ATP 0, 5 min, 30 min
Specification
VVEC
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Read More

Abcam user community

Verified customer

Submitted Jun 03 2016

Western blot abreview for Anti-PFKFB3 antibody [EPR12594]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (primary alveolar type II cells)
Gel Running Conditions
Non-reduced Denaturing (10%)
Loading amount
20 µg
Specification
primary alveolar type II cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More

Abcam user community

Verified customer

Submitted Nov 02 2015

Western blot abreview for Anti-PFKFB3 antibody [EPR12594]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (A549 cells)
Gel Running Conditions
Non-reduced Denaturing (10)
Loading amount
20 µg
Specification
A549 cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More

Dr. Christine Vohwinkel

Verified customer

Submitted Oct 29 2015

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