Product datasheet
Anti-Urm1 antibody ab4755
1 Abreviews 1 Image
Overview
Product name
Anti-Urm1 antibody
Description
Rabbit polyclonal to Urm1
Tested applications
WB, ELISA
Species reactivity
Reacts with: Saccharomyces cerevisiae
Immunogen
Recombinant full length protein (S. cerevisiae).
Positive control
Most yeast cell lysates can be used as a positive control without induction or stimulation.
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: 0.01% Sodium Azide
Constituents: 0.15M Sodium Chloride, 0.02M Potassium Phosphate. pH 7.2
Purity
IgG fraction
Purification notes
Purified from monospecific antiserum by a multi-step process which includes delipidation, salt
fractionation and ion exchange chromatography followed by extensive dialysis against the buffer
stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against antirabbit serum.
Clonality
Polyclonal
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab4755 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
1/1000. Detects a band of approximately 12 kDa.
ELISA
1/2000 - 1/10000.
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Target
Function
Acts as a sulfur carrier required for 2-thiolation of mcm(5)S(2)U at tRNA wobble positions of
tRNA(Lys), tRNA(Glu) and tRNA(Gln). Serves as sulfur donor in tRNA 2-thiolation reaction by
thiocarboxylated (-COSH) at its C-terminus by MOCS3. The sulfur is then transferred to tRNA to
form 2-thiolation of mcm(5)S(2)U. May also act as an ubiquitin-like protein that is covalently
conjugated to other proteins; the relevance of such function is however unclear in vivo.
Pathway
tRNA modification; 5-methoxycarbonylmethyl-2-thiouridine-tRNA biosynthesis.
Sequence similarities
Belongs to the URM1 family.
Post-translational
modifications
C-terminal thiocarboxylation occurs in 2 steps, it is first acyl-adenylated (-COAMP) via the
hesA/moeB/thiF part of MOCS3, then thiocarboxylated (-COSH) via the rhodanese domain of
MOCS3.
Cellular localization
Cytoplasm.
Anti-Urm1 antibody images
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ab4755 was tested by immunoblot against
yeast lysates expressing the Urm1-GFP
fusion protein and other UBL fusion proteins.
All UBLs possess limited homology to
Ubiquitin and to each other, therefore it is
important to know the degree of reactivity of
each antibody against each UBL. Panel A
shows total protein staining using ponceau.
Panel B shows positions of free GFP or GFP
containing recombinant proteins present in
each lysate preparation after reaction with a
Western blot - Urm1 antibody (ab4755)
1/1000 dilution of Goat polyclonal to GFP
(ab6673) followed by reaction with a 1/15000
dilution of Donkey polyclonal to Goat IgG
(HRP)(ab7125). Panel C shows specific
reaction with Urm1 using a 1/1000 dilution of
ab4755 followed by reaction with a 1/15000
dilution of Goat polyclonal to Rabbit IgG
(HRP) (ab7090). All primary antibodies were
diluted in TTBS buffer supplemented with 5%
non-fat milk and incubated with the
membranes overnight at 4° C. Yeast lysate
proteins were separated by SDS-PAGE
using a 15% gel. This data indicates that antiUrm1 is highly specific and does not cross
react with other UBLs. Bands present in
Panel C indicate that Urm1 and conjugated
Urm1 is present in most yeast cell lysates
albeit at significantly reduced levels relative to
the Urm1-GFP transfected lysate. A
chemiluminescence system was used for
signal detection (Roche). Other detection
systems will yield similar results. Data
contributed by M. Malakhov,
www.lifesensors.com, personal
communication.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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