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Rabbit Recombinant Monoclonal PFKFB3 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 73 publications.


Images

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFFlow Cyt (Intra)IHC-P
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Tested
Tested
Expected
Expected
Expected
Rat
Tested
Tested
Expected
Expected
Expected

Tested
Tested

Species

Human

Dilution info

1/50

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Mouse

Dilution info

1/50

Notes

-

Species

Rat

Dilution info

1/50

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000 - 1/10000

Notes

-

Species

Rat

Dilution info

1/1000 - 1/10000

Notes

-

Species

Mouse

Dilution info

1/1000 - 1/10000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/50

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Target data

Function

Synthesis and degradation of fructose 2,6-bisphosphate.

Alternative names

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Rabbit Recombinant Monoclonal PFKFB3 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 73 publications.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Clone number

EPR12594

Purification technique

Affinity purification Protein A

Concentration
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Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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 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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

Biological summary

PFKFB3 regulates cellular energy metabolism serving as a switch for converting carbohydrates into energy and building blocks for growth. This enzyme is part of a glycolytic complex coordinating energy needs during proliferation and survival. Its activity is important in processes demanding increased energy and anabolic precursors such as cell division. The enzyme's modulation affects fructose-26-bisphosphate levels influencing cellular responses to stress and nutrient availability directly impacting metabolic adaptation.

Mechanical summary

PFKFB3 also known as 6-phosphofructo-2-kinase/fructose-26-bisphosphatase 3 serves a significant role in regulating glycolysis. This bifunctional enzyme weighs approximately 58 kDa and manages levels of fructose-26-bisphosphate a potent activator of another enzyme called phosphofructokinase-1. PFKFB3 expression occurs in various tissues but is especially high in areas with active cell proliferation like the brain and tumors. This enzyme's activity affects glycolytic flux critical for energy production and biosynthesis especially under conditions requiring rapid energy supply.

Pathway

PFKFB3 integrates into the glycolytic pathway and the pentose phosphate pathway. PFKFB3's control over fructose-26-bisphosphate levels influences phosphofructokinase-1 modulating the glycolytic pathway's rate. This modulation impacts energy production efficiency and the provision of metabolites required for nucleotide synthesis linking to the pentose phosphate pathway. Proline-rich AKT substrate of 40 kDa (PRAS40) and AKT may also interact indirectly to regulate PFKFB3 activity in these metabolic pathways.

Disease

PFKFB3 links strongly to cancer and diabetes. Its upregulation in cancerous tissues supports tumor growth by promoting glycolysis known as the Warburg effect. Cancer-associated proteins such as KRAS often rely on PFKFB3's glycolytic control to sustain proliferative signaling. Additionally in diabetes altered PFKFB3 activity can contribute to insulin resistance due to disrupted metabolic homeostasis with insulin pathway proteins like IRS1 potentially interacting with it in this context.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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11 product images

  • Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861), expandable thumbnail

    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.

    All lanes: Western blot - Anti-PFKFB3 antibody [EPR12594] (AB181861)

    Predicted band size: 60 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-PFKFB3 antibody [EPR12594] (ab181861), expandable thumbnail

    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), expandable thumbnail

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

    Blocking buffer: 5% NFDM/TBST

    All lanes: Western blot - Anti-PFKFB3 antibody [EPR12594] (AB181861) at 1/20000 dilution

    Lane 1: Jurkat cell lysate at 20 µg

    Lane 2: HeLa cell lysate at 20 µg

    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

  • Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (ab181861), expandable thumbnail

    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.

    All lanes: Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (AB181861)

    Predicted band size: 60 kDa

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

    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 (Intracellular) - Anti-PFKFB3 antibody [EPR12594] (ab181861), expandable thumbnail

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

    Intracellular 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-PFKFB3 antibody [EPR12594] (ab181861), expandable thumbnail

    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).

  • Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (ab181861), expandable thumbnail

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

    PFKFB3 was immunoprecipitated from 0.35 mg of AR42J (rat pancreatic tumor epithelial cell) whole cell lysate with ab181861 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab181861 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

    Lane 1: AR42J (rat pancreatic tumor epithelial cell) whole cell lysate 10 μg (Input). 
    Lane 2: ab181861 IP in AR42J whole cell lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of  ab181861 in AR42J whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 180 seconds.

    All lanes: Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (AB181861)

    All lanes: Anti-PFKFB3 antibody IP in AR42J whole cell lysate

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (AB131366) at 1/5000 dilution

    Predicted band size: 60 kDa

    Observed band size: 60 kDa

    PFKFB3 was immunoprecipitated from 0.35 mg of AR42J (rat pancreatic tumor epithelial cell) whole cell lysate with ab181861 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab181861 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

    Lane 1: AR42J (rat pancreatic tumor epithelial cell) whole cell lysate 10 μg (Input). 
    Lane 2: ab181861 IP in AR42J whole cell lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of  ab181861 in AR42J whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 180 seconds.

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

  • Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (ab181861), expandable thumbnail

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

    PFKFB3 was immunoprecipitated from 0.35 mg of Mouse skin tissue lysate with ab181861 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab181861 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

    Lane 1: Mouse skin tissue lysate 10 μg (Input). 
    Lane 2: ab181861 IP in Mouse skin tissue lysate. 
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of  ab181861 in Mouse skin tissue lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 180 seconds.

    This blot was developed using a high sensitivity ECL substrate.

    All lanes: Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (AB181861)

    All lanes: Anti-PFKFB3 antibody in Mouse skin tissue lysate

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (AB131366) at 1/5000 dilution

    Observed band size: 60 kDa

    Exposure time: 180s

  • Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861), expandable thumbnail

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

    Exposure time: Lane 1-2: 26 seconds, Lane 3-7: 48 seconds

    Blocking buffer and concentration: 5% NFDM/TBST

    Lane 3-7 were developed using a high sensitivity ECL substrate.

    The expression level of PFKFB3 is upregulated during 3T3-L1 differentiation (PMID: 16306349).
    The band at approximately 110 kDa is likely to be PFKFB3 dimer (PMID: 31889092).

    All lanes: Western blot - Anti-PFKFB3 antibody [EPR12594] (AB181861) at 1/1000 dilution

    Lane 1: A431 (human epidermoid carcinoma epithelial cell), whole cell lysate at 20 µg

    Lane 2: bEnd.3 (mouse brain endothelial cell), whole cell lysate at 20 µg

    Lane 3: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

    Lane 4: 4T1 (mouse mammary gland carcinoma epithelial cell), whole cell lysate at 20 µg

    Lane 5: Undifferentiated 3T3-L1 (mouse embryonic fibroblast), whole cell lysate at 20 µg

    Lane 6: 3T3-L1 (mouse embryonic fibroblast) differentiated into adipocyte-like cells, whole cell lysate at 20 µg

    Lane 7: C2C12 (mouse myoblast), whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 60 kDa

    Observed band size: 60 kDa

  • Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861), expandable thumbnail

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

    Blocking buffer:  5% NFDM/TBST

    Exposure time: 15 seconds

    This blot was developed using a high sensitivity ECL substrate.

    All lanes: Western blot - Anti-PFKFB3 antibody [EPR12594] (AB181861) at 1/1000 dilution

    Lane 1: A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: Mouse skin tissue lysate at 20 µg

    Lane 3: Rat breast tissue lysate at 20 µg

    Lane 4: AR42J (rat pancreatic tumor epithelial cell) whole cell lysate at 20 µg

    Lane 5: L6 (rat skeletal muscle myoblast) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 60 kDa

    Observed band size: 60 kDa

    Exposure time: 15s

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Product protocols

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