|H M R Mk||Endogenous||52||Rabbit|
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.
SYAP1/BSTA Antibody recognizes endogenous levels of total SYAP1 protein.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys110 of human SYAP1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Synapse-associated protein 1 (SYAP1) was originally described by its similarity to the Drosophila synapse-associated SAP47 protein (1). Subsequent research using a yeast two-hybrid system described the protein as a BSD domain–containing signal transducer and Akt interactor (BSTA) based on protein structure and function. The ubiquitously expressed BSTA protein contains a central BSD domain that may play a role in mediating interaction between the BSTA protein and the serine/threonine kinase Akt1 (2). Research studies support a model of Akt1 activation that involves interactions between the BSTA protein and both Akt1 and the mTORC2 kinase complex, followed by phosphorylation of both BSTA and Akt1 by mTORC2. This series of interactions and phosphorylation events is thought to result in phosphorylation of Akt1 at Ser473 and Akt1 kinase activation (2). The BSTA mediated phosphorylation of Akt1 may promote adipocyte differentiation by suppressing the expression of the transcription factor FoxC2 (2). Additional studies show that the estrogen receptor antagonist tamoxifen can regulate the expression of BSTA in some breast cancer cells, suggesting a possible role for BSTA in pathways related to response to tamoxifen and other chemopreventative agents (3).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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|14077S||100 µl (10 western blots)||$ 255.0|