|H Mk||Endogenous||73,75||Rabbit IgG|
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 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: 263
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.
DCP1A (D6V1R) Rabbit mAb recognizes endogenous levels of total DCP1A protein. This antibody may also detect a nonspecific band of about 85 kDa. This antibody does not cross-react with DCP1B protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu436 of human DCP1A protein.
mRNA decapping is an important process in the mRNA turnover (1). DCP1A and DCP2 were identified as two human decapping enzymes and homologs of the better-characterized S. cerevisiae enzymes. Both putative decapping enzymes interact with the regulator of nonsense transcripts 1 (UPF1) and may be recruited by UPF1 or related proteins to mRNA sequences that contain premature termination codons (1). Additional research studies demonstrate that DCP1A, DCP1B (the homolog of DCP1A) and DCP2 colocalize with decapping activation factors RCK/p54 and Lsm proteins in cytoplasmic loci (2). DCP1A, DCP1B and DCP2 are components of cytoplasmic processing (P) bodies, with hyper-phosphorylation of DCP1A during mitosis suggesting a possible mechanism of P-body regulation during the cell cycle (3,4).
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|15365S||100 µl (10 western blots)||$ 255.0|