Product nameAnti-PFKFB3 antibody [EPR12594]
See all PFKFB3 primary antibodies
DescriptionRabbit monoclonal [EPR12594] to PFKFB3
Tested applicationsSuitable for: WB, IP, ICC/IF, IHC-P, Flow Cytmore details
Species reactivityReacts with: Mouse, Rat, Human
- Jurkat and HeLa whole cell lysate (ab150035); Human melanoma tissue; HeLa and A431 cells.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
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.
|WB||1/1000 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 60 kDa).|
|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.|
FunctionSynthesis and degradation of fructose 2,6-bisphosphate.
Sequence similaritiesIn the C-terminal section; belongs to the phosphoglycerate mutase family.
modificationsPhosphorylation by AMPK stimulates activity.
- Information by UniProt
- 6 phosphofructo 2 kinase/ fructose 2,6 bisphosphatase antibody
- 6 phosphofructo 2 kinase/fructose 2,6 biphosphatase 3 antibody
- 6-bisphosphatase antibody
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.
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).
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.
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
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 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.
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.
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).
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