Western blot analysis of extracts from various cell lines using NKG7 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
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
NKG7 Antibody recognizes endogenous levels of total NKG7 protein. This antibody may recognize multiple predicted isoforms of NKG7 protein.Species Reactivity:
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly161 of human NKG7 protein. Antibodies are purified by peptide affinity chromatography.
Natural Killer Granule Protein 7 (NKG7, also known as GMP-17 and GIG-I) is a novel type 1 integral membrane protein expressed at high levels in lymphocytes, including natural killer (NK) cells, γδ T cells, a subset of αβ T cells, lymphokine-activated killer (LAK) cells (1), and activated T regulatory cells (2). NKG7 expression can be induced in neutrophils following GM-CSF treatment (3). NKG7 protein is localized to the membranes of cytotoxic granules. Following target-cell induced NK cell degranulation, NKG7 translocates from granules to the plasma membrane (4). Studies have also shown that NKG7 is significantly up-regulated in allograft rejection patients (5-8).
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