使用敲除细胞株进行验证重组RabMAb

重组Anti-PFKFB3抗体[EPR12594] (ab181861)

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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: Mouse, Rat, Human

概述

  • 产品名称

    Anti-PFKFB3抗体[EPR12594]
    参阅全部 PFKFB3 一抗

  • 描述

    兔单克隆抗体[EPR12594] to PFKFB3

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, IP, ICC/IF, IHC-P, Flow Cytmore details

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Recombinant fragment within Human PFKFB3 aa 300 to the C-terminus. The exact sequence is proprietary.
    Database link: Q16875
    (Peptide available as ab192017)

  • 阳性对照

    • Jurkat and HeLa whole cell lysate (ab150035); Human melanoma tissue; HeLa and A431 cells.

  • 常规说明

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

性能

应用

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.

应用 Ab评论 说明
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. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.

靶标

  • 功能

    Synthesis and degradation of fructose 2,6-bisphosphate.

  • 组织特异性

    Ubiquitous.

  • 序列相似性

    In the C-terminal section; belongs to the phosphoglycerate mutase family.

  • 翻译后修饰

    Phosphorylation by AMPK stimulates activity.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • 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

图片

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

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

文献 (17)

发表研究结果有使用 ab181861？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab181861 被引用在 17 文献中.

  • 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
  • Pan H  et al. RNA binding protein HuR regulates extracellular matrix gene expression and pH homeostasis independent of controlling HIF-1a signaling in nucleus pulposus cells. Matrix Biol 77:23-40 (2019). PubMed: 30092282
  • Wang L  et al. Ablation of endothelial Pfkfb3 protects mice from acute lung injury in LPS-induced endotoxemia. Pharmacol Res 146:104292 (2019). PubMed: 31167111
View all Publications for this product

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1-4 of 4 Abreviews or Q&A

Western blot abreview for Anti-PFKFB3 antibody [EPR12594]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Murine breast cancer cells)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
5 µg
Specification
Murine breast cancer cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C
Read More

Alyssa La Belle

Verified customer

提交于 May 21 2019

Western blot abreview for Anti-PFKFB3 antibody [EPR12594]

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Jun 03 2016

Western blot abreview for Anti-PFKFB3 antibody [EPR12594]

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Nov 02 2015

Western blot abreview for Anti-PFKFB3 antibody [EPR12594]

 Excellent
Abreviews
abreview image
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
Read More

Dr. Christine Vohwinkel

Verified customer

提交于 Oct 29 2015

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