Name: Anti-PFKFB3 antibody [EPR12594]
Brand: RabMAb
SKU: ab181861
Rating: 5 (3 reviews)

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: Mouse, Rat, Human
Immunogen

Recombinant fragment within Human PFKFB3 aa 300 to the C-terminus. The exact sequence is proprietary.
Database link: Q16875

Run BLAST with Run BLAST with
Positive control
- Jurkat and HeLa whole cell lysate (ab150035); Human melanoma tissue; HeLa and A431 cells.
General 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 rabbit monoclonal antibody.

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

Alternative Versions
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 (ab102918)
- APC Conjugation Kit (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
Related Products
- Recombinant Human PFKFB3 protein (ab132993)
- Recombinant Human PFKFB3 protein (ab151856)

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		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.
Flow Cyt		Use at an assay dependent concentration.

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
- Unigene: 10791 Rat
see all
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

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)

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)

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)

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.
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)

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)

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).

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

This product has been referenced in:

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 . Read more (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). Read more (PubMed: 29113354) »

See all 4 Publications for this product

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

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1-3 of 3 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Jun 03 2016

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

Abcam user community

Verified customer

Submitted Nov 02 2015

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

Dr. Christine Vohwinkel

Verified customer

Submitted Oct 29 2015

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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Anti-PFKFB3 antibody [EPR12594] (ab181861)

Overview

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References

This product has been referenced in:

Customer reviews and Q&As

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