|H M R Mk||Endogenous||95||Rabbit IgG|
Western blot analysis of extracts from various cell lines using PATL1/PAT1b (D8P1B) Rabbit mAb.Learn more about how we get our images
Western blot analysis of extracts from 293T cells, mock transfected (-), transfected with a construct expressing DDK-tagged full-length human PATL1/PAT1b (hPATL1/PAT1b-DDK; +), or a contruct expressing HA-tagged full-length human PATL2 (hPATL2-HA; +), using PATL1/PAT1b (D8P1B) Rabbit mAb (upper), DYKDDDDK Tag (9A3) Mouse mAb #8146 (upper middle), HA-Tag (C29F4) Rabbit mAb #3724 (lower middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).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, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
PATL1/PAT1b (D8P1B) recognizes endogenous levels of total PATL1/PAT1b protein. This antibody does not cross-react with PATL2.
Human, Mouse, Rat, Monkey
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Tyr432 of human PATL1/PAT1b protein.
PATL1/PAT1b is the human homolog of the evolutionarily conserved Pat1/Mrt1 protein, which was first identified in Saccharomyces cerevisiae (1,2). This protein is a critical component of the RNA decay machinery in the cytoplasmic processing bodies (P-bodies), localized foci of mRNA silencing and degradation (3). PATL1/PAT1b interacts with many key components of the RNA decay machinery. These include the 5’ decapping proteins DDX6/RCK, DCP1, DCP2, EDC4, and the LSm1–7 ring (3,4) as well as the 3’ deadenylation complex of CAF1-CCR4-NOT1 (3).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. XP is a registered trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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|14288S||100 µl (10 western blots)||$ 255.0|