
Recombinant Anti-GFPT2 antibody [EPR19095] (ab190966)
- Datasheet
- References (2)
- Protocols
Overview
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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.
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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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19095 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
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 |
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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
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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
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Database links
- Entrez Gene: 9945 Human
- Entrez Gene: 14584 Mouse
- Entrez Gene: 360518 Rat
- Omim: 603865 Human
- SwissProt: O94808 Human
- SwissProt: Q9Z2Z9 Mouse
- SwissProt: Q4KMC4 Rat
- Unigene: 696497 Human
see all -
Alternative names
- D fructose 6 phosphate amidotransferase 2 antibody
- D-fructose-6-phosphate amidotransferase 2 antibody
- GFAT 2 antibody
see all
Images
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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®.
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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 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time:
Lane 1 & 2: 3 minutes.
Lane 3 & 4: 5 seconds.
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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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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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.
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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.
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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.
Protocols
Datasheets and documents
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) »