Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human IL-33 protein (hIL-33-Myc/DDK; +), using IL-33 (E2T5L) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from NCI-H1666 and JEG-3 cells using IL-33 (E2T5L) Rabbit mAb (upper) or 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 nonfat dry milk, 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.
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 June 2016
Protocol Id: 263
IL-33 (E2T5L) Rabbit mAb recognizes endogenous levels of total IL-33 protein.Species Reactivity:
Monoclonal antibody is produced by immunizing animals with human IL-33 recombinant protein and reacts with an epitope surrounding Glu165.
IL-33 (Interleukin-33, NF-HEV) is a nuclear cytokine that is part of the IL-1 family (1,2). Originally discovered as the ‘nuclear factor from high endothelial venules’, it is widely expressed by endothelial cells as well as epithelial barrier tissues (1,5). IL-33 is involved in regulating type 2-associated innate immune responses (2,4). It is the ligand for ST2, which along with IL-1RAcP, forms the IL-33R signaling complex (2,3). IL-33 functions as an alarmin upon cellular damage or stress, and is associated with allergic inflammation and asthma (5,6).
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.