|H M R Mk||Endogenous||50||Rabbit|
Western blot analysis of extracts from various cell lines using eRF1 Antibody.Learn more about how we get our images
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human eRF1 protein (heRF1-Myc/DDK; +), using eRF1 Antibody.Learn more about how we get our images
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody datasheet or product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised October 2016
Protocol Id: 10
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
eRF1 Antibody recognizes endogenous levels of total eRF1 protein.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Phe117 of human eRF1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Efficient termination of mRNA translation in eukaryotes is dependent upon a complex of two polypeptide release factors, eRF1 and eRF3 (1). The eukaryotic translation termination factor 1 (eRF1, ETF1) structurally resembles tRNA, which allows it to participate in stop codon recognition as well as hydrolysis of the peptidyl-tRNA conjugate (2,3). The eRF1 protein contains three functionally distinct domains, including an amino-terminal domain that harbors discrete motifs that participate in stop codon recognition (4,5). Lysine hydroxylation within the amino-terminal domain is required for efficient termination of mRNA translation (6). The central region of eRF1 harbors a GGQ motif that facilitates hydrolysis of peptidyl-tRNA conjugates (7), while its carboxy terminus participates in eRF3 binding (8).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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|13916S||100 µl (10 western blots)||$ 255.0|