Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with siRNA targeting human COPS6 (+), using COPS6 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from various cell lines using COPS6 Antibody.
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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
COPS6 Antibody recognizes endogenous levels of total COPS6 protein. This antibody also cross-reacts with an unidentifed protein of 78 kDa protein in some cell extracts.Species Reactivity:
Human, Mouse, Rat
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human COPS6 protein. Antibodies are purified by protein A and peptide affinity chromatography.
COP9 signalosome complex subunit 6 (COPS6) belongs to the multisubunit COP9 signalsome which modulates protein ubiquitination to affect cellular processes such as cell-cycle progression and DNA damage repair (1). COPS6 is a putative oncogene that has been shown to be overexpressed in a significant percentage of colorectal cancers with poor prognosis (2). Mechanistically, research studies have shown that a signaling axis involving EGFR and ERK promotes tumor growth through the phosphorylation of COPS6, which indirectly facilitates downstream stabilization of β-catenin and enhancement of its transcriptional acitivity (2). Reasearch studies have also demonstrated that COPS6 contributes to tumorigenesis through the stabilization of other oncoproteins like c-Myc, MDM2 and E6AP (3-5).
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