Western blot analysis of extracts from various cell lines using GRB7 (E7R8V) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Our Japan office is closed in observance of Emperor's Birthday. We will reopen on Tuesday, February 25th.
Thank you for your patience.
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
GRB7 (E7R8V) Rabbit mAb recognizes endogenous levels of total GRB7 protein.Species Reactivity:
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg29 of human GRB7 protein.
GRB7 belongs to the GRB7 family of adaptor proteins that mediate cell surface and cytosolic tyrosine kinases to downstream signaling pathways (1-3). The C-terminal SH2 domain of GRB7 has been shown to be important for its interaction with various upstream molecules such as ERBB receptor tyrosine kinases, FAK, EphB1, and SHC. The protein has an RA, CAM-BD, and PH domain located at the central region (GM region) of its sequences to mediate downstream Ras, PI3K/Akt, ERK, and Calmodulin signaling (3,4). Overexpression of GRB7 promotes cancer proliferation and migration, and disruption of GRB7 function inhibits its effect on cancer growth, invasion, and metastasis (3-5).
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.