|H||Endogenous||31, 33||Rabbit IgG|
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.
OTX2 (D7Y3J) Rabbit mAb recognizes endogenous levels of total OTX2 protein. This antibody recognizes both known human isoforms.
Mouse, Rat, Monkey, Chicken, Bovine, Guinea Pig, Horse
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asn251 of human OTX2 protein.
Orthodenticle homeobox 2 (OTX2) belongs to the bicoid subfamily of paired-box, homeodomain-containing transcription factors. OTX2 is a critically important neuronal transcription factor that functions to regulate the expression of cell cycle genes controlling proliferation and differentiation of neural progenitor cells (1-3). In addition to its neuronal development functions, it has been reported that OTX2 can function in a non-cell autonomous manner to promote survival of damaged retinal ganglion cells (4). OTX2 has also been shown to influence the susceptibility of post-mitotic neurons to toxic insult or physiological stress (3). Notably, aberrant expression of OTX2 has been strongly linked with neuronal tumor development. For example, research studies have found OTX2 is overexpressed in many medulloblastoma cell lines, and both overexpression and gene amplification were reported in a subset of primary medulloblastomas (5). In vitro studies support these observations, as targeted alterations in OTX2 expression directly affected both proliferation and senescence of medulloblastoma cell lines (6,7).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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|11943S||100 µl (10 western blots)||$ 255.0|