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RabMAb

Anti-PFKFB3抗体[EPR12594] (ab181861)

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
  • 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

  • 阳性对照
    • Jurkat and HeLa cell lysates; Human melanoma tissue; HeLa and A431 cells.
  • 常规说明

    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.

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.
Flow Cyt Use at an assay dependent concentration.

靶标

图片



  • Predicted band size : 60 kDa

    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.

  • 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).

  • 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
    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

  • 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 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.

  • 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.

     

  • 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).

文献

This product has been referenced in:
  • Jiang H  et al. PFKFB3-Driven Macrophage Glycolytic Metabolism Is a Crucial Component of Innate Antiviral Defense. J Immunol 197:2880-90 (2016). Read more (PubMed: 27566823) »
  • Lu Q  et al. Akt inhibition attenuates rasfonin-induced autophagy and apoptosis through the glycolytic pathway in renal cancer cells. Cell Death Dis 6:e2005 (2015). Human . Read more (PubMed: 26633711) »

See all 2 Publications for this product

客户评价及客户问答

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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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 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
Username

Dr. Christine Vohwinkel

Verified customer

提交于 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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