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

     

     

    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.

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

  • 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

  • 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®.

  • 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.

  • 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.

  • 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.

  • 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 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.

  • 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.

References

This product has been referenced in:

  • 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). Read more (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). Read more (PubMed: 30254013) »
See all 2 Publications for this product

Customer reviews and Q&As

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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