Western blot analysis of extracts from HT29 and GH3 cells, and GH3 cell conditioned medium (CM), using Shh (C9C5) Rabbit mAb.
|2207T||20 µl||$ 117|
|2207S||100 µl||$ 276|
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|MW (kDa)||19, (45 precursor)|
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.
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 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 June 2020
Protocol Id: 10
Shh (C9C5) Rabbit mAb detects the endogenous levels of total Shh protein. This antibody does not cross-react with transfected IHH and DHH.
Mouse, Chicken, Xenopus, Zebrafish
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu53 of human Shh.
Hedgehog proteins (Hh) are secreted signaling proteins that play many roles during animal development. Aberrant Hh signaling activity can be associated with numerous birth defects and uncontrolled Hh pathway activation is linked to the development of several types of cancers (1-2). The three identified vertebrate Hh genes are Sonic (Shh), Indian (Ihh) and Desert (Dhh), all of which have distinct as well as overlapping roles (3-5). Hh proteins are synthesized as 45 kDa precursors that undergo auto-cleavage to generate a 19 kDa amino-terminal peptide (Hh-N) and a carboxy-terminal peptide (Hh-C). The amino-terminal peptide becomes covalently attached to a cholesterol molecule at its carboxy terminus and acetylated at its amino terminus. This doubly modified Hh-N peptide is released from cells and responsible for all known Hedgehog signaling activity (6).
Explore pathways + proteins related to this product.
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