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.
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: 262
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.
ZFX (L28B6) Mouse mAb detects endogenous levels of total ZFX protein.
Monoclonal antibody is produced by immunizing animals with a recombinant fragment of human ZFX.
The ZFX (X-linked zinc finger protein) gene is expressed from the inactive X chromosome and is structurally similar to its homologue on the Y chromosome, ZFY (1). Transcripts of ZFX and ZFY encode proteins composed of a highly acidic amino-terminal domain and a carboxy-terminal zinc-finger motif that is commonly associated with nucleic acid-binding proteins (2). Both ZFY and ZFX are probable transcriptional activators and may function in sex determination (3). Multiple alternatively spliced transcript variants of ZFX, encoding different isoforms, have been identified and may be functionally distinct (2). Conditional gene targeting studies in mouse have suggested ZFX is also required for self-renewal of embryonic and hematopoietic stem cells (4). ZFX has also been suggested to play a role in proliferation and expansion of B cells, and could contribute to lymphocyte homeostasis (5).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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|5419S||100 µl (10 western blots)||$255.00.0|