Western blot analysis of extracts from U-937, THP-1, and HuH-6 cells, untreated (-) or treated with PNGase F (+), using Cathepsin G Antibody (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The lack of detectable Cathepsin G expression in HuH-6 cells is consistent with proteomic and transcriptomic expression profiling data, confirming specificity of the antibody.
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.
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
Cathepsin G Antibody recognizes endogenous levels of total Cathepsin G protein.Species Reactivity:
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro130 of human Cathepsin G protein. Antibodies are purified by peptide affinity chromatography.
Cathepsin G is a serine protease which was first identified in the cytoplasmic azurophilic granules of neutrophil leukocytes (1). As a degradative enzyme, Cathepsin G acts both intracellularly to degrade ingested host pathogens and extracellularly to breakdown ECM components at inflammatory sites (2). Independent of its serine protease activity, Cathepsin G is also known to exert broad-spectrum antibacterial action against Gram-negative and –positive bacteria (3). Cathepsin G has been reported to play an important role in the pathophysiology of several diseases, such as chronic obstructive pulmonary disease (COPD), rheumatoid arthritis, cystic fibrosis and other conditions clinically manifested by excessive inflammatory reactions (4-7). Because of its role in host defense and disease, Cathepsin G is of interest as a potential therapeutic target (8,9).
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