Recombinant Anti-PFKFB3 antibody [EPR12594] (ab181861)
![Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861)](https://www.abcam.com/ps/products/181/ab181861/Images/ab181861-286155-anti-pfkfb3-antibody-epr12594-western-blot.jpg "Western blot - 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
Overview
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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
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Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Jurkat and HeLa whole cell lysate (ab150035); Human melanoma tissue; HeLa and A431 cells.
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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
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
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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
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
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).
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| IP |
1/50.
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| ICC/IF |
1/100.
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| IHC-P |
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| Flow Cyt |
Use at an assay dependent concentration.
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| Notes |
|---|
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WB
1/1000 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 60 kDa). |
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IP
1/50. |
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ICC/IF
1/100. |
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IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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Flow Cyt
Use at an assay dependent concentration. |
Target
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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
modificationsPhosphorylation by AMPK stimulates activity. - Information by UniProt
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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
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Alternative names
- 6 phosphofructo 2 kinase/ fructose 2,6 bisphosphatase antibody
- 6 phosphofructo 2 kinase/fructose 2,6 biphosphatase 3 antibody
- 6-bisphosphatase antibody
see all
Images
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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.
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![Immunocytochemistry/ Immunofluorescence - Anti-PFKFB3 antibody [EPR12594] (ab181861)](https://www.abcam.com/ps/products/181/ab181861/Images/ab181861-216608-anti-pfkfb3-antibody-epr12594-immunofluorescence.jpg)
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).
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![Western blot - Anti-PFKFB3 antibody [EPR12594] (ab181861)](https://www.abcam.com/ps/products/181/ab181861/Images/ab181861-216600-anti-pfkfb3-antibody-epr12594-western-blot.jpg)
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
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PFKFB3 antibody [EPR12594] (ab181861)](https://www.abcam.com/ps/products/181/ab181861/Images/ab181861-216604-anti-pfkfb3-antibody-epr12594-immunohistochemistry.jpg)
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.
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![Flow Cytometry - Anti-PFKFB3 antibody [EPR12594] (ab181861)](https://www.abcam.com/ps/products/181/ab181861/Images/ab181861-268995-anti-pfkfb3-antibody-epr12594-flow-cytometry.jpg)
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.
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![Immunoprecipitation - Anti-PFKFB3 antibody [EPR12594] (ab181861)](https://www.abcam.com/ps/products/181/ab181861/Images/ab181861-216611-anti-pfkfb3-antibody-epr12594-immunoprecipitation.jpg)
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.
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![Immunocytochemistry/ Immunofluorescence - Anti-PFKFB3 antibody [EPR12594] (ab181861)](https://www.abcam.com/ps/products/181/ab181861/Images/ab181861-216610-anti-pfkfb3-antibody-epr12594-immunofluorescence.jpg)
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).
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![Anti-PFKFB3 antibody [EPR12594] (ab181861)](https://www.abcam.com/ps/products/181/ab181861/Images/ab181861-8-benefits-of-recombinant-antibodies.png)
Anti-PFKFB3 antibody [EPR12594] (ab181861)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (20)
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