Western blot analysis of extracts from various cell lines using CSDE1 Antibody (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
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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
CSDE1 Antibody recognizes endogenous levels of total CSDE1 protein.Species Reactivity:
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human CSDE1 protein. Antibodies are purified by peptide affinity chromatography.
Originally described as the product of an active transcription unit located upstream of the N-RAS gene (1), CSDE1 (cold shock domain-containing protein E1) is a highly conserved RNA-binding protein containing five cold-shock domains (2,3). CSDE1 is mostly localized in the cytoplasm where it interacts with distinct protein complexes involved in the post-transcriptional regulation of numerous mRNAs in a dynamic and complex manner (4). CSDE1 plays a role in maintaining the undifferentiated state of human embryonic stem cells (hESCs), and its loss results in accelerated neural differentiation and neurogenesis, partially via post-transcriptional regulation of fatty acid binding protein 7 (FABP7) and vimentin (VIM) mRNAs (5). In human melanoma, increased CSDE1 protein expression has been shown to modulate the levels of pro-oncogenic factors such as vimentin or RAC1 and tumor suppressors such as PTEN, thus promoting tumor invasion and metastasis (6).
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