|H M R Mk||Endogenous||43||Rabbit|
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 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: 263
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.
CCN3 Antibody recognizes endogenous levels of total CCN3 protein.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg118 of human CCN3 protein. Antibodies are purified by protein A and peptide affinity chromatography.
CCN3, also named NOV (Nephroblastoma overexressed), belongs to the CCN (Cyr61, Ctgf, NOV) family of proteins. It is a cystine-rich secretory protein that associates with components of the extracellular matrix. Like other CCN family members, CCN3 is capable of mediating diverse biological functions through its four distinct domains, which enable binding to numerous protein partners (1-5).
CCN3 modulates bone turnover through various mechanisms and is implicated in the progression of primary bone cancers such as osteosarcoma and chondrosarcoma (6-8). Research has shown that CCN3 is also involved in the bone metastasis of melanoma, breast cancer, and prostate cancers (9-11). Recently, CCN3 was reported to play an important role in stem cell renewal (12). CCN3 is normally expressed in both embryonic and adult tissues (13,14). The activity of CCN3 is influenced by post translational modifications and proteolytic cleavage (15,16).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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|8767S||100 µl||$ 260.0|