|H M R Mk||Endogenous||68, 71||Rabbit|
Western blot analysis of extracts from various cell lines using NPL4 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 NPL4 protein (hNPL4-Myc/DDK; +), using NPL4 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.
NPL4 Antibody recognizes endogenous levels of total NPL4 protein.
Human, Mouse, Rat, Monkey
Chicken, Xenopus, Bovine, Dog, Horse
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys322 of human NPL4 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Nuclear protein localization protein 4 (NPL4, NPLOC4) was originally identified as a yeast nuclear transport protein that was later recognized as a critical component of the endoplasmic reticulum-associated degradation (ERAD) pathway (1,2). Mammalian NPL4 protein has an amino-terminal ubiquitin-like domain containing a p97 binding site, and a conserved carboxy-terminal zinc finger (NZF) motif responsible for binding ubiquitinated target proteins (2,3). NPL4 binds ubiquitin fusion degradation protein 1 (UFD1) to form a heterodimer that associates with the p97 AAA-ATPase, creating a protein complex that mediates delivery of ubiquitinated ER proteins to the proteasome (4,5).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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|13489S||100 µl (10 western blots)||$ 255.0|