|H M R Mk||Endogenous||51||Mouse IgG1|
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.
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 June 2016
Protocol Id: 262
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
ZPR1 (LG1) Mouse mAb detects endogenous levels of total ZPR1 protein.
Human, Mouse, Rat, Monkey
Monoclonal antibody is produced by immunizing animals with purified recombinant human ZPR1.
The zinc finger protein ZPR1 (ZNF259) binds to epidermal growth factor receptor (EGFR) and is localized to both cytoplasm and nucleus. The zinc fingers found in ZPR1 and the tyrosine kinase domain of EGFR mediate the interaction between ZPR1 and the receptor (1). ZPR1 translocates from the cytoplasm to nucleus following mitogen (i.e. EGF) stimulation (2,3). ZPR1 also interacts with translation elongation factor eEF1A in vivo following EGF treatment (3). The interaction between the zinc finger protein and elongation factor is important for cell proliferation. Cells lacking ZPR1 exhibit abnormal nucleolar function, suggesting that ZPR1 is required for cell viability and nucleolar function in dividing cells (3). ZPR1 knockout mice exhibit significant neurodegeneration, and reduced or altered expression of ZPR1 may contribute to spinal muscular atrophy, a disorder characterized by degeneration of spinal cord neurons (4).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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