RecombinantRabMAb

Recombinant Anti-GFPT2 antibody [EPR19095] (ab190966)

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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 [EPR19095] to GFPT2
  • Suitable for: WB, ICC/IF, IP, Flow Cyt
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-GFPT2 antibody [EPR19095]
    See all GFPT2 primary antibodies

  • Description

    Rabbit monoclonal [EPR19095] to GFPT2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, IP, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human GFPT2 aa 1-300. The exact sequence is proprietary.
    Database link: O94808

  • Positive control

    • WB: Human GFPT2 recombinant protein fragment; Human placenta and glioma lysates; Mouse placenta, heart, kidney and spleen lysates; Rat placenta, heart and spleen lysates; A-375, U-87MG and NIH/3T3 whole cell lysates. ICC/IF: U-87 MG cell line. Flow Cyt: U-87 MG cell line. IP: NIH/3T3 whole cell lysate.

  • 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
    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab190966 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 1/2000. Detects a band of approximately 77 kDa (predicted molecular weight: 77 kDa).
ICC/IF 1/500.
IP 1/50.
Flow Cyt 1/150.

Target

  • Function

    Controls the flux of glucose into the hexosamine pathway. Most likely involved in regulating the availability of precursors for N- and O-linked glycosylation of proteins.

  • Tissue specificity

    Highest levels of expression in heart, placenta, and spinal cord.

  • Pathway

    Nucleotide-sugar biosynthesis; UDP-N-acetyl-alpha-D-glucosamine biosynthesis; alpha-D-glucosamine 6-phosphate from D-fructose 6-phosphate: step 1/1.

  • Sequence similarities

    Contains 1 glutamine amidotransferase type-2 domain.
    Contains 2 SIS domains.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • D fructose 6 phosphate amidotransferase 2 antibody
    • D-fructose-6-phosphate amidotransferase 2 antibody
    • GFAT 2 antibody
    • GFAT2 antibody
    • Gfpt2 antibody
    • GFPT2_HUMAN antibody
    • Glucosamine fructose 6 phosphate aminotransferase [isomerizing] 2 antibody
    • glutamine fructose 6 phosphate transaminase 2 antibody
    • Glutamine--fructose-6-phosphate aminotransferase [isomerizing] 2 antibody
    • Glutamine:fructose 6 phosphate amidotransferase 2 antibody
    • Glutamine:fructose-6-phosphate amidotransferase 2 antibody
    • Hexosephosphate aminotransferase 2 antibody
    see all

Images

  • Western blot - Anti-GFPT2 antibody [EPR19095] (ab190966)
    Western blot - Anti-GFPT2 antibody [EPR19095] (ab190966)
    All lanes : Anti-GFPT2 antibody [EPR19095] (ab190966) at 1/2000 dilution

    Lane 1 : Human GFPT2 recombinant protein fragment
    Lane 2 : Human GFPT1 recombinant protein fragment

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 77 kDa
    Observed band size: 33 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.
    Exposure time: Lane 1: 1 second; Lane 2: 3 minutes.

    Human GFPT2 recombinant protein fragment contains aa2-305 with a His-Tag®, Human GFPT1 recombinant protein fragment contains aa2-288 with a His-Tag®.

  • Western blot - Anti-GFPT2 antibody [EPR19095] (ab190966)
    Western blot - Anti-GFPT2 antibody [EPR19095] (ab190966)
    All lanes : Anti-GFPT2 antibody [EPR19095] (ab190966) at 1/2000 dilution

    Lane 1 : Human placenta lysate
    Lane 2 : Mouse placenta lysate
    Lane 3 : A-375 (Human malignant melanoma cell line) whole cell lysate
    Lane 4 : Rat placenta lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 77 kDa
    Observed band size: 77 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 

    Lane 1 & 2: 3 minutes.

    Lane 3 & 4: 5 seconds.

  • Western blot - Anti-GFPT2 antibody [EPR19095] (ab190966)
    Western blot - Anti-GFPT2 antibody [EPR19095] (ab190966)
    All lanes : Anti-GFPT2 antibody [EPR19095] (ab190966) at 1/2000 dilution

    Lane 1 : Mouse heart lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat heart lysate
    Lane 5 : Rat spleen lysate
    Lane 6 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 77 kDa
    Observed band size: 77 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-GFPT2 antibody [EPR19095] (ab190966)
    Western blot - Anti-GFPT2 antibody [EPR19095] (ab190966)
    All lanes : Anti-GFPT2 antibody [EPR19095] (ab190966) at 1/2000 dilution

    Lane 1 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate
    Lane 2 : Human glioma lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lane 2 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 77 kDa
    Observed band size: 77 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-GFPT2 antibody [EPR19095] (ab190966)
    Immunocytochemistry/ Immunofluorescence - Anti-GFPT2 antibody [EPR19095] (ab190966)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling GFPT2 with ab190966 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasm staining on U-87 MG cell line. 

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab190966 at 1/500 dilution followed by ab150120 at 1/1000 dilution.

    -ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • Flow Cytometry - Anti-GFPT2 antibody [EPR19095] (ab190966)
    Flow Cytometry - Anti-GFPT2 antibody [EPR19095] (ab190966)

    Flow cytometric analysis of 4% paraformaldehyde-fixed U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling GFPT2 with ab190966 at 1/150 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue).

    Goat anti-Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-GFPT2 antibody [EPR19095] (ab190966)
    Immunoprecipitation - Anti-GFPT2 antibody [EPR19095] (ab190966)

    GFPT2 was immunoprecipitated from 1mg of NIH/3T3 (Mouse embryo fibroblast cells) whole cell lysate with ab190966 at 1/50 dilution.

    Western blot was performed from the immunoprecipitate using ab190966 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: NIH/3T3 whole cell lysate 10ug (Input).

    Lane 2: ab190966 IP in NIH/3T3 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab190966 in NIH/3T3 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 5 seconds.

  • Anti-GFPT2 antibody [EPR19095] (ab190966)
    Anti-GFPT2 antibody [EPR19095] (ab190966)

References (2)

Publishing research using ab190966? Please let us know so that we can cite the reference in this datasheet.

ab190966 has been referenced in 2 publications.

  • Shi GD  et al. iTRAQ-based proteomics profiling of Schwann cells before and after peripheral nerve injury. Iran J Basic Med Sci 21:832-841 (2018). PubMed: 30186571
  • Dai W  et al. Consumption of a high fat diet promotes protein O-GlcNAcylation in mouse retina via NR4A1-dependent GFAT2 expression. Biochim Biophys Acta Mol Basis Dis 1864:3568-3576 (2018). PubMed: 30254013

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Western blot abreview for Anti-GFPT2 antibody [EPR19095]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
HeLa
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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Verified customer

Submitted Jan 12 2018

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