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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PFKFB3 antibody [EPR12594] (ab181861)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (4) Submit a question References (20)

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Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861)
  • Immunocytochemistry/ Immunofluorescence - Anti-PFKFB3 antibody [EPR12594] (ab181861)
  • Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PFKFB3 antibody [EPR12594] (ab181861)
  • Flow Cytometry - Anti-PFKFB3 antibody [EPR12594] (ab181861)
  • Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (ab181861)
  • Immunocytochemistry/ Immunofluorescence - Anti-PFKFB3 antibody [EPR12594] (ab181861)
  • Anti-PFKFB3 antibody [EPR12594] (ab181861)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR12594] to PFKFB3
  • Suitable for: WB, IP, ICC/IF, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human

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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: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped 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 buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR12594
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Hypoxia
    • Hypoxia-Regulated
    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Neuroscience
    • Neurology process
    • Metabolism
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-PFKFB3 antibody [EPR12594] (ab203651)
    • Alexa Fluor® 488 Anti-PFKFB3 antibody [EPR12594] (ab203984)
    • Anti-PFKFB3 antibody [EPR12594] - BSA and Azide free (ab218121)
    • Anti-Indoleamine 2, 3-dioxygenase antibody [EPR1230Y] - Low endotoxin, Azide free (ab224262)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab30128)
    • Jurkat whole cell lysate (ab7899)
  • Recombinant Protein

    • Recombinant Human PFKFB3 protein (ab151856)
  • Related Products

    • Recombinant Human PFKFB3 protein (ab151856)

Applications

The Abpromise guarantee

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 (4)
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.
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.
  • Target information above from: UniProt accession Q16875 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5209 Human
    • Entrez Gene: 170768 Mouse
    • Entrez Gene: 117276 Rat
    • Omim: 605319 Human
    • SwissProt: Q16875 Human
    • SwissProt: A2AUP1 Mouse
    • SwissProt: O35552 Rat
    • Unigene: 195471 Human
    • 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).

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

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (20)

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

    ab181861 has been referenced in 20 publications.

    • Piers TM  et al. A locked immunometabolic switch underlies TREM2 R47H loss of function in human iPSC-derived microglia. FASEB J 34:2436-2450 (2020). PubMed: 31907987
    • Liu J  et al. Long noncoding RNA AGPG regulates PFKFB3-mediated tumor glycolytic reprogramming. Nat Commun 11:1507 (2020). PubMed: 32198345
    • Zhang J  et al. Dual inhibition of PFKFB3 and VEGF normalizes tumor vasculature, reduces lactate production, and improves chemotherapy in glioblastoma: insights from protein expression profiling and MRI. Theranostics 10:7245-7259 (2020). PubMed: 32641990
    • Li D  et al. Variation in energy metabolism arising from the effect of the tumor microenvironment on cell biological behaviors of bladder cancer cells and endothelial cells. Biofactors 46:64-75 (2020). PubMed: 31580525
    • Hu X  et al. PI3K-Akt-mTOR/PFKFB3 pathway mediated lung fibroblast aerobic glycolysis and collagen synthesis in lipopolysaccharide-induced pulmonary fibrosis. Lab Invest 100:801-811 (2020). PubMed: 32051533
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-4 of 4 Abreviews or Q&A

    Western blot abreview for Anti-PFKFB3 antibody [EPR12594]

    Excellent
    Abreviews
    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
    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
    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
    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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