Western blot analysis of recombinant human CXCL10 protein using CXCL10 (D5L5L) Rabbit mAb.
Western blot analysis of extracts from HT-29 cells, untreated (-) or treated with Human Interferon-γ (hIFN-γ) #8901 (50 ng/ml, 16 hr; +), using CXCL10 (D5L5L) Rabbit mAb (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, 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
CXCL10 (D5L5L) Rabbit mAb recognizes endogenous levels of total CXCL10 protein. This antibody also cross-reacts with a 100kDa protein of unknown orgin in some cell lines.Species Reactivity:
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to human CXCL10 protein.
Chemokine (C-X-C motif) ligand 10 (CXCL10, IP-10) is a chemokine produced by a number of cell types and that is induced in response to pro-inflammatory cytokines, such as IFN-γ and TNF-α (1,2). CXCL10 binds to the chemokine receptor CXCR3 and induces calcium flux and cell migration (3). The CXCR3 protein is normally expressed on activated CD4+ T cells and NK cells, which can produce pro-inflammatory cytokines, including IFN-γ and TNF-α (3,4). As a result, these molecules act in an amplification loop to promote immune responses in the contexts of both autoimmunity and a tumor immune response (5,6).
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