Western blot analysis of extracts from either 293T cells, mock transfected (-) or transfected (+) with a cDNA expression construct encoding full-length human EGR3 (hEGR3), or mouse and rat brain tissue using EGR3 Antibody.
|REACTIVITY||H M R|
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.
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
EGR3 Antibody recognizes endogenous levels of total EGR3 protein.Species Reactivity:
Human, Mouse, Rat
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Phe160 of human EGR3 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Early growth response 3 (EGR3) is a zinc finger transcription factor and one of four members of the early growth response family (1). Members of this family are basally expressed in the brain (2) and are immediate early response genes that are important for the induction of cellular programs of differentiation, proliferation, and cell death in response to environmental stimuli (1). EGR3 plays important roles in cellular growth and neuronal development (2) and an essential role in learning and memory processing of both short- and long-term hippocampus-dependent memory; it also mediates adaptation to stress and novelty (3). Research studies show that a lack of EGR3 results in abnormal neuronal and T cell development (4).
Explore pathways + proteins related to this product.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.