WB
Western blot analysis of PFKFB3 on different lysates with Rabbit anti-PFKFB3 antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: PC-12 cell lysate Lane 3: A431 cell lysate Lane 4: A549 cell lysate Lysates/proteins at 20 µg/Lane. Exposure time: 2 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-PFKFB3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
ICC staining of PFKFB3 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).IP
PFKFB3 was immunoprecipitated from 0.2 mg HeLa cell lysate with PFKFB3 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using PFKFB3 antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: PFKFB3 antibody IP in HeLa cell lysate Lane 3: Rabbit IgG instead of PFKFB3 antibody in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 2 minutesFC
Flow cytometric analysis of PFKFB3 was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | PFKFB3 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within Human PFKFB3 aa 260-520 / 520. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:500-1:1000 IHC:1:1000 ICC:1:50-1:200 IP:Use at an assay dependent concentration. FC:1:50-1:100 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | PFKFB3 |
|---|---|
| Gene Synonyms | PFK2 IPFK2 iPFK-2 |
| Gene Full Name | 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 |
| Gene Summary | The protein encoded by this gene belongs to a family of bifunctional proteins that are involved in both the synthesis and degradation of fructose-2,6-bisphosphate, a regulatory molecule that controls glycolysis in eukaryotes. The encoded protein has a 6-phosphofructo-2-kinase activity that catalyzes the synthesis of fructose-2,6-bisphosphate (F2,6BP), and a fructose-2,6-biphosphatase activity that catalyzes the degradation of F2,6BP. This protein is required for cell cycle progression and prevention of apoptosis. It functions as a regulator of cyclin-dependent kinase 1, linking glucose metabolism to cell proliferation and survival in tumor cells. Several alternatively spliced transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Apr 2016] |
| 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 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 2 antibody |
| Molecular Weight(MW) | 60kDa |
| Cellular Localization | Cytosol. Nucleoplasm. |
WB
Western blot analysis of PFKFB3 on different lysates with Rabbit anti-PFKFB3 antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: PC-12 cell lysate Lane 3: A431 cell lysate Lane 4: A549 cell lysate Lysates/proteins at 20 µg/Lane. Exposure time: 2 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue using anti-PFKFB3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
ICC staining of PFKFB3 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
IP
PFKFB3 was immunoprecipitated from 0.2 mg HeLa cell lysate with PFKFB3 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using PFKFB3 antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: PFKFB3 antibody IP in HeLa cell lysate Lane 3: Rabbit IgG instead of PFKFB3 antibody in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 2 minutes
FC
Flow cytometric analysis of PFKFB3 was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:500-1:1000 IHC:1:1000 ICC:1:50-1:200 IP:Use at an assay dependent concentration. FC:1:50-1:100 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
