Product datasheet
Anti-Phosphoserine/threonine antibody ab17464
9 Abreviews 10 References 5 Images
Overview
Product name
Anti-Phosphoserine/threonine antibody
Description
Rabbit polyclonal to Phosphoserine/threonine
Specificity
Detects phosphoserine or phosphothreonine in the context of tyrosine, tryptophan or
phenylalanine at the -1 position or phenylalanine at the +1 position. The antibody does not
recognize the nonphosphorylated form of these motifs, nor does it recognize other
phosphoserine or phosphothreonine containing proteins and peptides.
Tested applications
ICC/IF, ELISA, IP, WB
Species reactivity
Reacts with: Species independent
Immunogen
Synthetic peptide.
Positive control
Calyculin A treated A431 cells
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer
Preservative: None
Constituents: 50% Glycerol, BSA, 150mM Sodium chloride, 10mM HEPES. pH 7.5
Purity
Protein A purified
Clonality
Polyclonal
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab17464 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
ICC/IF
1/100.
ELISA
1/1000.
IP
1/100.
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Application
Abreviews
WB
Notes
1/1000.
Block with 5% nonfat milk, 0.1% Tween-20. Dilute primary antibody in 5% BSA,
0.1% Tween-20.
Target
Relevance
A hallmark of signal transduction pathways is the reversible phosphorylation of serine and
threonine residues within specific sequences, or motifs, in target proteins. Specific signaling
motifs include not only sequences that are recognized by protein kinases, but also those that are
recognized by phosphorylation-dependent binding proteins like 14-3-3. These modular
phosphoprotein interacting domains are critical elements in modulating, directing and amplifying
intracellular communications. Many critical protein kinases can be regulated by phosphorylation
at a specific serine or threonine surrounded by phenylalanine or tyrosine. For example, Akt, an
important kinase that regulates cell survival, is activated by phosphorylation at Ser473, a site
surrounded by phenylalanine and tyrosine. RSK1, p70 S6 K, and certain PKC isoforms also
contain a similar consensus phosphorylation site. Phosphorylation of these sites is required for
kinase activity. The Phospho-(Ser/Thr) Phe Antibody is a powerful tool for discovery of new
proteins containing this important regulatory motif.
Anti-Phosphoserine/threonine antibody images
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All lanes : Anti-Phosphoserine/threonine
antibody (ab17464) at 1/500 dilution
Lane 1 : Whole tissue lysate prepared from
murine heart (control)
Lane 2 : Whole tissue lysate prepared from
murine heart (control)
Lane 3 : Whole tissue lysate prepared from
murine heart (PKC)
Western blot - Phosphoserine/threonine antibody
Lane 4 : Whole tissue lysate prepared from
(ab17464)
murine heart (PKC)
Image courtesy of an anonymous Abreview.
Lysates/proteins at 50 µg per lane.
Secondary
HRP conjugated goat anti-rabbit polyclonal at
1/5000 dilution
developed using the ECL technique
Observed band size : 150 kDa
Exposure time : 5 minutes
Image courtesy of an anonymous Abreview.
The lower blot is to detect total mybpc3. It was
probed with a non-abcam anti-mybpc3
antibody.
ab17464 staining Phosphoserine/threonine in
murine cardiomyocytes by
Immunocytochemistry/ Immunofluorescence.
Cells were fixed in paraformaldehyde,
permeabilized using 1% Triton X-100,
Immunocytochemistry/ Immunofluorescence -
blocked with 10% horse serum for 1 hour at
Anti-Phosphoserine/threonine antibody (ab17464)
room temperature and then incubated with
Image courtesy of an anonymous Abreview.
ab17464 at a 1/100 dilution for 2 hours. The
secondary used was an Alexa-Fluor 488
conjugated goat anti-rabbit polyclonal used at
a 1/500 dilution.
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Image from Gao X et al, PLoS Pathog
5:e1000708 (2009), Fig S3.
To determine if NleH could phosphorylate
RPS3, we conducted in vitro kinase assays
with NleH and RPR3 and analyzed the results
by Immunoblotting with ab17464, phosphoSer/Thr-specific antibody, following
Western blot - Phosphoserine/threonine antibody
separation by SDS-PAGE.
(ab17464)
Image from Gao X et al, PLoS Pathog 5:e1000708
(2009), Fig S3.
All lanes : Anti-Phosphoserine/threonine
antibody (ab17464) at 1/1000 dilution
Lane 1 : Human THP-1 Whole Cell Extract
Lane 2 : Mouse Total Lung Extract
Lysates/proteins at 10 µg per lane.
Secondary
Alkaline phosphatase conjugated goat antirabbit antibody
Western blot - Phosphoserine/threonine antibody
developed using the ECL technique
(ab17464)
This image is courtesy of an anonymous Abreview
Performed under reducing conditions.
Exposure time : 5 minutes
This image is courtesy of an anonymous
Abreview
All lanes : Anti-Phosphoserine/threonine
antibody (ab17464) at 1/1000 dilution
Lane 1 : calyculin A treated A431 cells
Lane 2 : calyculin A treated A431 cells
Western blot - Phosphoserine/threonine antibody
(ab17464)
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