|H M R Mk||Endogenous||60||Rabbit IgG|
Western blot analysis of extracts from various cell lines using PFKFB3 (D7H4Q) Rabbit mAb.Learn more about how we get our images.
Western blot analysis of extracts from 293 cells, mock transfected (-) or transfected with a construct expressing DDK-tagged full-length human PFKFB3 (hPFKFB3-DDK; +), using PFKFB3 (D7H4Q) Rabbit mAb (upper), DYKDDDDK Tag (9A3) Mouse mAb #8146 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).Learn more about how we get our images.
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
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.
PFKFB3 (D7H4Q) Rabbit mAb recognizes endogenous levels of total PFKFB3 protein.
Human, Mouse, Rat, Monkey
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu456 of human PFKFB3 protein.
The bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2/FBPase or PFKFB) catalyzes the synthesis and degradation of fructose 2,6-bisphosphate and regulates its steady-state level (1,2). Fructose 2,6-bisphosphate activates phosphofructokinase, a rate-limiting enzyme in glycolysis, by allosteric regulation (1,2). Four different PFKFB isoforms (PFKFB1, PFKFB2, PFKFB3, and PFKFB4) have been identified (1,2). One of them, PFKFB3/iPFK-2, was shown to be inducible by hypoxia leading to increased glycolysis under hypoxic conditions (2). Research studies have shown that PFKFB3/iPFK-2 is also highly expressed in some types of human cancer (3).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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|13123S||100 µl (10 western blots)||$ 255.0|