Product datasheet
Anti-PAG3 (phospho Y724) antibody ab18031
1 Image
Overview
Product name
Anti-PAG3 (phospho Y724) antibody
Description
Rabbit polyclonal to PAG3 (phospho Y724)
Specificity
Lysates prepared from COS cells overexpressing human wild-type PAG3 and cotransfected with
mouse Pyk2 were immunoblotted in the presence of non-phosphopeptide corresponding to the
immunogen, a generic phosphotyrosine-containing peptide or the phosphopeptide immunogen.
The data show that only the peptide corresponding to phospho Y724 PAG3 blocks the signal,
and the signal is absent when mutant PAG3 extracts are used, demonstrating the specificity of
the antibody.
Tested applications
WB
Species reactivity
Reacts with human PAG3 Predicted to react with mouse PAG3 due to 92% sequence homology
Immunogen
Synthetic phosphopeptide derived from the region of human PAG3 that contains tyrosine 724.
Positive control
COS cells overexpressing human wild type PAG3, cotransfected with mouse Pyk2.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or 80°C. Avoid freeze / thaw cycle.
Storage buffer
Preservative: 0.05% Sodium Azide
Constituents: 50% Glycerol, PBS (without Mg2+ and
Ca2+), 1mg/ml BSA. pH 7.3.
Purity
Immunogen affinity purified
Purification notes
This antibody was purified from rabbit serum by sequential epitope specific chromatography.
The antibody has been negatively preadsorbed using a non phosphopeptide corresponding to
the site of phosphorylation to remove antibody that is reactive with non phosphorylated PAG3
protein. The final product is generated by affinity chromatography using a PAG3 derived peptide
that is phosphorylated at tyrosine 724.
Clonality
Polyclonal
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab18031 in the following tested applications.
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The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Abreviews
WB
Notes
1/1000. Detects a band of approximately 160 kDa (predicted molecular weight:
112 kDa).
Target
Function
Activates the small GTPases ARF1, ARF5 and ARF6. Regulates the formation of post-Golgi
vesicles and modulates constitutive secretion. Modulates phagocytosis mediated by Fc gamma
receptor and ARF6. Modulates PXN recruitment to focal contacts and cell migration.
Tissue specificity
Detected in heart, brain, placenta, kidney, monocytes and pancreas.
Sequence similarities
Contains 2 ANK repeats.
Contains 1 Arf-GAP domain.
Contains 1 PH domain.
Contains 1 SH3 domain.
Domain
The conserved Arg-464 in the Arf-GAP domain probably becomes part of the active site of
bound small GTPases and is necessary for GTP hydrolysis.
Post-translational
modifications
Phosphorylated on tyrosine residues by SRC and PTK2B.
Cellular localization
Cytoplasm. Golgi apparatus > Golgi stack membrane. Cell membrane. Colocalizes with F-actin
and ARF6 in phagocytic cups.
Anti-PAG3 (phospho Y724) antibody images
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Predicted band size : 112 kDa
Western blot using Ab18031 on COS cell
lysate overexpressing human PAG3.
Western blot - PAG3 (phospho Y724) antibody
(ab18031)
Lane 1: wild-type PAG3 alone
Lane 2: wild-type PAG3 cotransfected with
mouse PyK2
Lane 3: wild-type PAG3 cotransfected with
mouse PyK2. Antibody blocked with the non
phosphopeptde corresponding to the
immunogen
Lane 4: wild-type PAG3 cotransfected with
mouse PyK2. Antibody blocked with the
generic phosphotyrosine corresponding to the
immunogen
Lane 5: wild-type PAG3 cotransfected with
mouse PyK2. Antibody blocked with the
generic phosphopeptide immunogen
Lane 6: mutant [Y724/Y763] PAG3
cotransfected with Pyk2
10-30 micrograms of cell lysate can be
loaded when using similar lysates with this
antibody. Samples were run using SDSPAGE on a 8% polyacrylamide gel and
transferred to PVDF. Membranes were
blocked with a 5% BSA-TBST buffer for one
hour at room temperature, then incubated with
PAG3 antibody Ab18031 for two hours at
room temperature in a 3% BSA-TBST buffer,
following prior incubation with blocking
peptides. After washing, membranes were
incubated with goat F(ab’)2 anti-rabbit IgG
HRP conjugate in 3% BSA-TBST buffer, and
bands were detected using the Pierce
SuperSignal method.
Only the phospho peptide immunogen blocks
the antibody, confirming its specificity for the
phospho epitope tyrosine 724.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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