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
NOTE: Do not allow slides to dry at any time during this procedure.
For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; follow with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.
posted June 2005
revised March 2016
Protocol Id: 303
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.
TCL1 Antibody detects endogenous levels of total TCL1.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the amino-terminal sequence of human TCL1. Antibodies are purified by protein A and peptide affinity chromatography.
TCL1 (T cell leukemia 1), MTCP1 and TCL1b belong to the TCL1 proto-oncogene family, and their products are involved in Akt activation during embryonic development, T cell leukemias, prolymphocytic leukemias and B cell lymphomas (1-3). The Akt association domain of TCL1 binds with the PH domain of Akt. The formation of an oligomeric TCL-Akt complex is required for TCL1 coactivator function and results in phosphorylation and activation of Akt . Furthermore, functional analysis indicates that the interaction between TCL1 and Akt promotes translocation of Akt to the nucleus (4-6). These findings are supported by the crystal structure of TCL1, which suggests that TCL1 may participate in molecular transport (7).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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|4042S||100 µl (10 western blots)||$ 255.0|