Western blot analysis of extracts from HCC827 cells, mock transfected or transfected with GFP, using GFP (D5.1) XP® Rabbit mAb (HRP Conjugate).
Supplied in 136 mM NaCl, 2.6 mM KCI, 12 mM sodium phosphate (pH 7.4) dibasic, 2 mg/ml BSA, and 50% glycerol. Store at –20°C. Do not aliquot the antibodies.
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.
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: 264
GFP (D5.1) XP® Rabbit mAb (HRP Conjugate) detects GFP-tagged proteins exogenously expressed in cells.
All Species Expected
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of GFP. This antibody was conjugated to HRP under optimal conditions.
Epitope tags are useful for the labeling and detection of proteins using immunoblotting, immunoprecipitation, and immunostaining techniques. Because of their small size, they are unlikely to affect the tagged protein’s biochemical properties.
The green-fluorescent protein (GFP) functions as a bioluminescence energy transfer acceptor in the jellyfish Aequorea that maximally absorbs light at 395 nm and has an emission spectrum that peaks at 509 nm. GFP has become a very useful tool as a fusion protein that reports gene expression, traces cell lineage and defines subcellular protein localization (1).
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XP is a registered trademark of Cell Signaling Technology, Inc.