Western blot analysis of extracts from various cell lines using CdGAP (D6J9G) Rabbit mAb.
|REACTIVITY||H M R Mk|
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 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
CdGAP (D6J9G) Rabbit mAb recognizes endogenous levels of total CdGAP protein.Species Reactivity:
Human, Mouse, Rat, Monkey
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser520 of human CdGAP protein.
The Rho family of small GTPases, including Rho, Rac, and Cdc42, act as molecular switches that regulate processes such as cell migration, adhesion, proliferation, and differentiation. They are activated by guanine nucleotide exchange factors (GEFs), which catalyze the exchange of bound GDP for GTP, and inhibited by GTPase activating proteins (GAPs), which catalyze the hydrolysis of GTP to GDP (1). The serine- and proline-rich GAP protein, Cdc42 GAP (CdGAP), has been shown to be a negative regulator of both Cdc42 and Rac1, but not RhoA (2,3). This protein contains three domains: an amino-terminal GAP domain, a central domain, and a carboxy-terminal proline-rich domain containing five Src homology 3 (SH3)-binding sites. It is suggested that threonine and serine phosphorylation within the proline-rich domain likely alters protein-protein interactions and determines the localization of CdGAP (4). Phosphorylation of CdGAP on threonine 776 by both ERK-1 and GSK-3 has been shown to negatively regulate protein activity, possibly by inducing a conformational change within the protein disrupting its ability to bind SH3 domains (4,5). Upregulation of CdGAP has been shown to increase cell proliferation and it has been suggested that this protein may play a role in TGF-β-induced cell growth, motility, and invasion in some breast cancer cells (6).
Explore pathways + proteins related to this product.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Tween is a registered trademark of ICI Americas, Inc.