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

Rabbit Recombinant Monoclonal PFKFB3 antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 5 publications.

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Images

Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] - BSA and Azide free (AB218121), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

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

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Tested	Tested	Not recommended	Expected	Not recommended
Rat	Tested	Tested	Not recommended	Expected	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info -	Notes -
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human, Rat, Mouse	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat, Mouse	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat, Mouse	Dilution info -	Notes -

Associated Products

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2 products for Alternative Version

3 products for Alternative Product

Target data

See full target information PFKFB3

Function

Catalyzes both the synthesis and degradation of fructose 2,6-bisphosphate.

Alternative names

PFK/FBPase 3, Renal carcinoma antigen NY-REN-56, iPFK-2, PFKFB3

Recommended products

Rabbit Recombinant Monoclonal PFKFB3 antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 5 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR12594
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab218121 is the carrier-free version of Anti-PFKFB3 antibody [EPR12594] ab181861.

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.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

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.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity 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.

Biological function 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.

Pathways

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.

Associated diseases and disorders

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

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10 product images

Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] - BSA and Azide free (ab218121)
Western blot analysis of PFKFB3 in HeLa cell lysate immunoprecipitated using Anti-PFKFB3 antibody [EPR12594] 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 (Anti-PFKFB3 antibody [EPR12594] ab181861).
All lanes: Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (Anti-PFKFB3 antibody [EPR12594] ab181861)
Predicted band size: 60 kDa
Flow Cytometry (Intracellular) - Anti-PFKFB3 antibody [EPR12594] - BSA and Azide free (ab218121)
Intracellular Flow Cytometry analysis of A431 (human epidermoid carcinoma) cells labeling PFKFB3 with purified Anti-PFKFB3 antibody [EPR12594] 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 (Anti-PFKFB3 antibody [EPR12594] ab181861).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PFKFB3 antibody [EPR12594] - BSA and Azide free (ab218121)
This IHC data was generated using the same anti-PFKFB3 antibody clone, EPR12594, in a different buffer formulation (cat# Anti-PFKFB3 antibody [EPR12594] ab181861).
Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling PFKFB3 with Anti-PFKFB3 antibody [EPR12594] 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.
Immunocytochemistry/ Immunofluorescence - Anti-PFKFB3 antibody [EPR12594] - BSA and Azide free (ab218121)
Immunofluorescent analysis of acetone-fixed HeLa cells labeling PFKFB3 with Anti-PFKFB3 antibody [EPR12594] 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 (Anti-PFKFB3 antibody [EPR12594] ab181861).
Immunocytochemistry/ Immunofluorescence - Anti-PFKFB3 antibody [EPR12594] - BSA and Azide free (ab218121)
Immunofluorescent analysis of 4% paraformaldehyde-fixed A431 cells labeling PFKFB3 with Anti-PFKFB3 antibody [EPR12594] 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 (Anti-PFKFB3 antibody [EPR12594] ab181861).
Western blot - Anti-PFKFB3 antibody [EPR12594] - BSA and Azide free (ab218121)
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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PFKFB3 antibody [EPR12594] ab181861).
All lanes: Western blot - Anti-PFKFB3 antibody [EPR12594] (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) (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
Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] - BSA and Azide free (ab218121)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PFKFB3 antibody [EPR12594] ab181861).
PFKFB3 was immunoprecipitated from 0.35 mg of Mouse skin tissue lysate with Anti-PFKFB3 antibody [EPR12594] ab181861 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-PFKFB3 antibody [EPR12594] ab181861 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (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: Anti-PFKFB3 antibody [EPR12594] ab181861 at 1/30 IP in Mouse skin tissue lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-PFKFB3 antibody [EPR12594] ab181861 in Mouse skin tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST
This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.
All lanes: Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (Anti-PFKFB3 antibody [EPR12594] ab181861) at 1/1000 dilution
Lane 1: Mouse skin tissue lysate at 10 µg
Lane 2: Anti-PFKFB3 antibody [EPR12594] ab181861 at 1/30 IP in Mouse skin tissue lysate at 1/30 dilution
Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 in Mouse skin tissue lysate at 10 µg
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Developed using the ECL technique.
Observed band size: 60 kDa
Exposure time: 180s
Western blot - Anti-PFKFB3 antibody [EPR12594] - BSA and Azide free (ab218121)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PFKFB3 antibody [EPR12594] ab181861).
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.
All lanes: Western blot - Anti-PFKFB3 antibody [EPR12594] (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) (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
Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] - BSA and Azide free (ab218121)
PFKFB3 was immunoprecipitated from 0.35 mg of AR42J (rat pancreatic tumor epithelial cell) whole cell lysate with Anti-PFKFB3 antibody [EPR12594] ab181861 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-PFKFB3 antibody [EPR12594] ab181861 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (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: Anti-PFKFB3 antibody [EPR12594] ab181861 IP in AR42J whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-PFKFB3 antibody [EPR12594] 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 (Anti-PFKFB3 antibody [EPR12594] ab181861).
All lanes: Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (Anti-PFKFB3 antibody [EPR12594] ab181861)
Lane 1: AR42J (rat pancreatic tumor epithelial cell) whole cell lysate at 10 µg
Lane 2: Anti-PFKFB3 antibody IP in AR42J whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Predicted band size: 60 kDa
Observed band size: 60 kDa
Western blot - Anti-PFKFB3 antibody [EPR12594] - BSA and Azide free (ab218121)
PFKFB3 Western blot staining using rabbit Anti-PFKFB3 antibody
This data was developed using Anti-PFKFB3 antibody [EPR12594] ab181861, the same antibody clone in a different buffer formulation.
Low expression: kidney tissue (PMID: 34660743).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/1000000 dilution.
All lanes: Western blot - Anti-PFKFB3 antibody [EPR12594] (Anti-PFKFB3 antibody [EPR12594] ab181861) at 1/1000 dilution
Lane 1: Mouse lung tissue lysate at 20 µg
Lane 2: Mouse liver tissue lysate at 20 µg
Lane 3: Mouse heart tissue lysate at 20 µg
Lane 4: Mouse kidney tissue lysate at 20 µg
Lane 5: Rat lung tissue lysate at 20 µg
Lane 6: Rat liver tissue lysate at 20 µg
Lane 7: Rat heart tissue lysate at 20 µg
Lane 8: Rat kidney tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Developed using the ECL technique.
Observed band size: 60 kDa
Exposure time: 180s